Figures
Figure 1 with 2 supplements
Kupffer cell (KC) death is a characteristic feature of metabolic dysfunction-associated steatotic liver disease (MASLD) progression.
(A–D) Male wild-type C57BL/6J mice were fed a high-fat high-cholesterol diet (HFHC) for 0, 4, or 16 weeks. (A) KC death was assessed by immunostaining of liver sections for TIM4 (KC marker, red), TdT-mediated dUTP Nick-End Labeling (TUNEL) (green), and DAPI (nuclei, blue). Scale bar: 50 µm (main panels) and 20 µm (inset). (B) KC death was quantified. n=4 mice/group. (C) Flow cytometry analysis of KCs (CD45+ F4/80hi CD11blow TIM4+) and infiltrating macrophages (IMs) (CD45+ F4/80lowCD11bhi TIM4-) among isolated nonparenchymal cells (NPCs). (D) KC counts were quantified. n=4–5 mice/group. (E–F) Male wild-type C57BL/6J mice were fed either: (E) normal chow diet (NCD) or high-fat diet (HFD) for 20 weeks, or (F) NCD or methionine-choline-deficient diet (MCD) for 6 weeks. KC death was assessed by immunostaining of liver sections for TIM4 (green), TUNEL (red), and DAPI (nuclei, blue). Scale bar: 50 µm (main panels) and 20 µm (inset). KC death was quantified. n=4 mice/group. Representative images are shown in A, C, E, F. One-way ANOVA (B, D). Unpaired Student’s t-test (E, F). p-Value as indicated.
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Figure 1—source data 1
Numerical data of Figure 1B–D and E–F.
- https://cdn.elifesciences.org/articles/109206/elife-109206-fig1-data1-v1.xlsx
Figure 1—figure supplement 1
The generation of high-fat high-cholesterol diet (HFHC)-induced metabolic dysfunction-associated steatotic liver disease (MASLD) mouse model.
Male wild-type C57B/6J mice were fed with HFHC for 0, 4, or 16 weeks. (A) Hematoxylin and eosin (H&E) (top), Oil Red O (middle), and Sirius Red staining (bottom) were performed to detect MASLD progression in liver sections at 0, 4, or 16 weeks after HFHC, respectively. Scale bar: 20 μm. Liver fibrosis was quantified. MASLD activity score is diagnosed. (B) Body weight was recorded during HFHC feeding. (C) Serum ALT, AST, cholesterol, triglyceride, or liver cholesterol triglyceride is measured at 16 weeks after HFHC. n=4 mice/group. Unpaired Student’s t-test (B, C); one-way ANOVA (A). p-Value as indicated. ns: not significant.
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Figure 1—figure supplement 1—source data 1
Numerical data of Figure 1—figure supplement 1A–C.
- https://cdn.elifesciences.org/articles/109206/elife-109206-fig1-figsupp1-data1-v1.xlsx
Figure 1—figure supplement 2
Examination of Kupffer cell (KC) death and monocyte-derived macrophages (MoMFs) recruitment in high-fat high-cholesterol diet (HFHC) mice.
Male wild-type (WT) C57B/6J mice were fed with HFHC for 0, 4, or 16 weeks. (A) KC death was examined by immunofluorescence staining of Clec4f and cleavaged caspase 3 (Cl-Casp3) and DAPI (nuclei) in livers of mice. KC death was quantified. n=4 mice/group. (B) Flow cytometry analysis of MoMFs (CD45+Ly6G⁻CD11b+F4/80lowTIM4low/-) among nonparenchymal cells (NPCs) of WT mice fed HFHC diet for 0, 4, or 16 weeks. (C) MoMFs counts were quantified. n=4–5 mice/group. (D) Proliferation-associated TdT-mediated dUTP Nick-End Labeling (TUNEL+) KCs was examined by co-staining of Ki67, TIM4, TUNEL, and DAPI (nuclei). Ki67+ KCs among TUNEL+ KCs was quantified. Representative images are shown in A, D. One-way ANOVA (A, C, D). p-Value as indicated. Scale bar: 20 μm (main panels), 5 μm (inset).
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Figure 1—figure supplement 2—source data 1
Numerical data of Figure 1—figure supplement 2A and C–D.
- https://cdn.elifesciences.org/articles/109206/elife-109206-fig1-figsupp2-data1-v1.xlsx
Figure 2
Kupffer cells (KCs) exhibit early and zone-specific susceptibility to death during metabolic dysfunction-associated steatotic liver disease (MASLD) progression.
(A–E) Male wild-type C57BL/6J mice were fed a high-fat high-cholesterol diet (HFHC) for 0, 4, or 16 weeks. (A–C) Hepatic cell death was assessed by co-staining TdT-mediated dUTP Nick-End Labeling (TUNEL) with: (A) HNF4α (hepatocytes), (B) Desmin (hepatic stellate cells [HSCs]), (C) Iba1 (hepatic macrophages), and DAPI (nuclei, blue). Scale bars: 50 µm (main panels). Hepatic cell death was quantified (n=4 mice/group). (D) Zonal distribution of KC death was evaluated by co-staining TIM4 (KCs), TUNEL, glutamine synthetase (GS, central vein marker), and DAPI (nuclei, blue). Scale bars: 50 µm. Zonal distribution of KC death was quantified (n=6–7 mice/group). FOV: field of view. PV: portal vein. CV: central vein. Representative images are shown in A–D. One-way ANOVA (A–D). p-Value as indicated.
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Figure 2—source data 1
Numerical data of Figure 2A–D.
- https://cdn.elifesciences.org/articles/109206/elife-109206-fig2-data1-v1.xlsx
Figure 3 with 1 supplement
Kupffer cells (KCs) exhibit metabolic reprogramming with increased glycolysis during early metabolic dysfunction-associated steatotic liver disease (MASLD).
(A) Experimental design for metabolomic analysis of KCs isolated from male wild-type mice fed a high-fat high-cholesterol diet (HFHC) for 0, 4, or 8 weeks. n=3 mice/group. (B) Principal component analysis (PCA) of enriched metabolites in KCs across different dietary durations. (C–D) Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis of metabolic pathways upregulated in KCs at 4 weeks (C) or 8 weeks (D). The glucose metabolism pathway is highlighted by red rectangles. (E) Heatmap depicting significantly altered metabolites involved in glucose metabolism pathways in KCs across different dietary durations. (F) Heatmap depicting significantly altered metabolites involved in cell death in KCs across different dietary durations.
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Figure 3—source data 1
Numerical data of Figure 3B–F.
- https://cdn.elifesciences.org/articles/109206/elife-109206-fig3-data1-v1.xlsx
Figure 3—figure supplement 1
Dynamic changes in mRNA expression levels of rate-limiting enzyme genes involved in glucose metabolism.
(A) Purity of isolated Kupffer cells (KCs) was examined by immunofluorescence staining of TIM4 and DAPI (nuclei). Scale bar: 20 μm (main panels), 5 μm (inset). Purity was quantified (n=6 independent experiments). (B) qRT-PCR analysis of mRNA expression levels of key rate-limiting enzymes in glycolysis, pentose phosphate pathway (PPP), glycogenolysis, glycogenesis, TCA cycle, and β-oxidation in wild-type (WT) KCs from mice for indicated dietary durations (n=4 mice/group). One-way ANOVA (B), p-values as indicated.
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Figure 3—figure supplement 1—source data 1
Numerical data of Figure 3—figure supplement 1A–B.
- https://cdn.elifesciences.org/articles/109206/elife-109206-fig3-figsupp1-data1-v1.xlsx
Figure 4 with 1 supplement
Excessive glucose metabolic activity contributes to Kupffer cell (KC) death.
(A–B) Isolated KCs were treated for 24 hr with: 5.5 mM glucose+isopropanol (control), 5.5 mM glucose+800 µM palmitic acid (PA), 10 mM glucose+800 µM PA. Cell viability was assessed by Cleaved caspase 3 (Cl-Casp3) staining (Cl-Casp3+ cells = dead). Scale bars: 20 µm (main panels), 5 µm (insets). Cl-Casp3 was detected by western blot. (C–D) Isolated KCs were treated for 24 hr with: blank (no treatment), DMSO (vehicle control), 20 µM PS48 (PDK1 activator). Scale bars: 20 µm (main panels), 5 µm (insets). Cell death was analyzed as above. (E) Experimental design. Male wild-type (WT) mice were fed a high-fat high-cholesterol diet (HFHC) for 5 weeks. From the third week onward, mice received intraperitoneal injections of either vehicle or 2-DG (50 mg/kg) every other day (n=3–4 mice/group). (F–G) Effects of glycolysis inhibition on KC death after 5 weeks of HFHC feeding. (F) Representative images of liver sections co-stained with TdT-mediated dUTP Nick-End Labeling (TUNEL) and the KC marker TIM4. (G) Quantification of TUNEL+ KCs. Data are presented as mean ± SEM. Statistical analysis was performed using one-way ANOVA (A, C) and an unpaired Student’s t-test (G); p-values are indicated.
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Figure 4—source data 1
Numerical data of Figure 4A, C and G.
- https://cdn.elifesciences.org/articles/109206/elife-109206-fig4-data1-v1.xlsx
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Figure 4—source data 2
PDF file containing original western blots for Figure 4B and D, indicating the relevant bands and treatments.
- https://cdn.elifesciences.org/articles/109206/elife-109206-fig4-data2-v1.pdf
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Figure 4—source data 3
Original files for western blot analysis displayed in Figure 4B and D.
- https://cdn.elifesciences.org/articles/109206/elife-109206-fig4-data3-v1.zip
Figure 4—figure supplement 1
Excessive glucose metabolic activity contributes to Kupffer cell (KC) death.
(A) Calcein-AM was used to assess primary KC viability treated as in main Figure 4. Scale bar: 20 μm. Representative images are shown. (B–C) Isolated KCs were treated for 24 hr with: blank (no treatment), DMSO (vehicle control), 20 µM oligomycin (Oligo, ATP synthase inhibitor). Scale bars: 20 µm (main panels), 5 µm (insets). Cell death was analyzed as above. One-way ANOVA (B). p-Value as indicated.
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Figure 4—figure supplement 1—source data 1
Numerical data of Figure 4—figure supplement 1B.
- https://cdn.elifesciences.org/articles/109206/elife-109206-fig4-figsupp1-data1-v1.xlsx
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Figure 4—figure supplement 1—source data 2
PDF file containing original western blots for Figure 4—figure supplement 1C, indicating the relevant bands and treatments.
- https://cdn.elifesciences.org/articles/109206/elife-109206-fig4-figsupp1-data2-v1.pdf
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Figure 4—figure supplement 1—source data 3
Original files for western blot analysis displayed in Figure 4—figure supplement 1C.
- https://cdn.elifesciences.org/articles/109206/elife-109206-fig4-figsupp1-data3-v1.zip
Figure 5 with 2 supplements
Enhanced glycolytic flux in Chi3l1-/- macrophages.
(A) Schematic diagram depicting the fate of glucose-derived ribose carbons in wild-type (WT) mouse macrophages. (B) Principal component analysis (PCA) of metabolites in WT and Chi3l1-/- bone marrow-derived macrophages (BMDMs) cultured with [U-13C]glucose. (C) Heatmap depicting significantly altered glycolysis and pentose phosphate (PPP) metabolites in WT and Chi3l1-/- BMDMs. (D) Glucose metabolic flux analysis in WT and Chi3l1-/- BMDMs cultured with [U-13C]glucose showing mass isotopologue distributions of: glycolytic intermediates (Glc, F6P, FBP, 3PGA, 2PGA, PEP, PA, LA, G6P) and PPP intermediates (Ru5P, R5P, S7P, DHAP). Data represent n=6 biological replicates/group. (E–F) Extracellular acidification rate (ECAR) analysis of WT or Chi3l1-/- BMDM cells. BMDMs were sequentially treated with glucose, oligomycin, and 2-DG as indicated during seahorse. Unpaired Student’s t-test (D, F). *p<0.05, **p<0.01, ***p<0.001.
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Figure 5—source data 1
Numerical data of Figure 5B–F.
- https://cdn.elifesciences.org/articles/109206/elife-109206-fig5-data1-v1.xlsx
Figure 5—figure supplement 1
Comparison of Kupffer cell (KC) polarization between wild-type (WT) and Chi3l1-/- mice during metabolic dysfunction-associated steatotic liver disease (MASLD) progression.
(A) qPCR analysis of key genes involved in macrophage polarization pathway in liver tissues of WT and Chi3l1-/- mice fed with high-fat high-cholesterol diet (HFHC) at indicated week. n=4–7 mice/group. Representative images are shown in A. One-way ANOVA (A). p-Value as indicated.
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Figure 5—figure supplement 1—source data 1
Numerical data of Figure 5—figure supplement 1A.
- https://cdn.elifesciences.org/articles/109206/elife-109206-fig5-figsupp1-data1-v1.xlsx
Figure 5—figure supplement 2
Recombinant Chi3l1 (rChi3l1) inhibits glucose utilization in Chi3l1-/- bone marrow-derived macrophages (BMDMs).
(A) Comparative principal component analysis (PCA) of metabolites in Chi3l1-/- and Chi3l1-/- supplemented with rChi3l1 BMDMs cultured with uniformly labeled [U-13C]glucose. (B) Heatmap depicting significantly altered glycolysis and pentose phosphate pathway (PPP) metabolites in Chi3l1-/- and Chi3l1-/- supplemented with rChi3l1 BMDMs. (C) Glucose metabolic flux analysis in Chi3l1-/- and Chi3l1-/- supplemented with rChi3l1 BMDMs showing mass isotopologue distributions of: glycolysis pathway intermediates (Glc, F6P, FBP, 3PGA, 2PGA, PEP, PA, LA, G6P) and PPP intermediates (Ru5P, R5P, S7P, DHAP). Data represent n=6 biological replicates. (D) Lactate dehydrogenase (LDH) activity in culture medium of Chi3l1-/- BMDMs treated for 24 hr with: 10 mM glucose (high glucose), 10 mM glucose+100 ng/mL rChi3l1. Unpaired Student’s t-test (D). p-Value as indicated.
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Figure 5—figure supplement 2—source data 1
Numerical data of Figure 5—figure supplement 2A–D.
- https://cdn.elifesciences.org/articles/109206/elife-109206-fig5-figsupp2-data1-v1.xlsx
Figure 6 with 2 supplements
Enhanced glycolysis accelerated Kupffer cell (KC) death during metabolic dysfunction-associated steatotic liver disease (MASLD).
(A) Cleaved caspase 3 (Cl-Casp3) staining to detect wild-type (WT) and Chi3l1-/- KC death. Cells were under treatment without (blank) or with either isopropyl alcohol (Iso) or palmitic acid (PA) for 24 hr. Scale bar: 20 μm. (B) Cl-Casp3+ cells were quantified. (C) Lactate dehydrogenase (LDH) release measurement in culture medium of KCs isolated from male WT and Chi3l1-/- mice was measured after treatment for 24 hr with: blank (no treatment), ISO (vehicle control), 800 µM PA. (D) Flow cytometry analysis of KCs (CD45+ F4/80hi CD11blow TIM4+) and monocyte-derived macrophages (MoMFs) (CD45+ F4/80low CD11bhi TIM4-) among nonparenchymal cells (NPCs) in Clec4f-cre and Chi3l1-KpKO mice fed high-fat high-cholesterol diet (HFHC) for 0 or 16 weeks. (E) KC counts were quantified. n=4 mice/group. (F) KC death was assessed by immunostaining of TIM4 (KC marker, green), TdT-mediated dUTP Nick-End Labeling (TUNEL) (red), and DAPI (nuclei, blue) in liver sections from Clec4f-cre and Chi3l1-KpKO mice fed HFHC for 0 or 16 weeks. Scale bar: 50 μm (main panels) and 20 μm (inset). (G) KC death was quantified. n=4 mice/group. Representative images shown (A, D, F). Unpaired Student’s t-test (B, C, E, G). p-Value as indicated.
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Figure 6—source data 1
Numerical data of Figure 6B–C, E and G.
- https://cdn.elifesciences.org/articles/109206/elife-109206-fig6-data1-v1.xlsx
Figure 6—figure supplement 1
Chi3l1 is majorly expressed in hepatic macrophages, and its expression is upregulated during metabolic dysfunction-associated steatotic liver disease (MASLD).
(A) Cellular source of Chi3l1 was assessed by immunohistochemical analysis of consecutive liver sections from mice fed a high-fat high-cholesterol diet (HFHC) for 16 weeks. Serial sections were independently stained for Chi3l1 and lineage markers: HNF4α (hepatocytes), Desmin (hepatic stellate cells [HSCs]), or Iba1 (hepatic macrophages). Cellular localization was determined by aligning morphological profiles across sequential sections. Scale bars: 20 μm (main panels), 5 μm (insets). Hepatocytes are outlined in Chi3l1+ images in green dashed lines. (B) Chi3l1 expression in KCs was assessed by co-staining with F4/80 (red) and TIM4 (white) in livers of mice fed either normal chow diet (NCD, 0 weeks) or HFHC (16 weeks). Scale bar: 20 μm (main panels), 5 μm (inset). The intensity of Chi3l1 expression in KCs was quantified. Unpaired Student’s t-test (B). p-Value as indicated.
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Figure 6—figure supplement 1—source data 1
Numerical data of Figure 6—figure supplement 1B.
- https://cdn.elifesciences.org/articles/109206/elife-109206-fig6-figsupp1-data1-v1.xlsx
Figure 6—figure supplement 2
Cre insertion promotes Kupffer cell (KC) self-proliferation.
(A) Comparision of KC self-proliferation in wild-type (WT) or Clec4f cre mice under normal chow diet (NCD) or Clec4f cre mice under high-fat high-cholesterol diet (HFHC) feeding by co-staining of Ki67 (proliferation marker) and TIM4 (KC marker). Nuclei are counterstained with DAPI (blue). Scale bar: 20 μm (main panels) and 10 μm (inset). (B) KC proliferation was quantified. Representative images are shown in A. One-way ANOVA (B). p-Value as indicated.
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Figure 6—figure supplement 2—source data 1
Numerical data of Figure 6—figure supplement 2B.
- https://cdn.elifesciences.org/articles/109206/elife-109206-fig6-figsupp2-data1-v1.xlsx
Figure 7
Excessive glycolysis enhancement promotes Kupffer cell (KC) death in metabolic dysfunction-associated steatotic liver disease (MASLD).
(Left) Under physiological conditions, KCs maintain basal glucose metabolism supporting cellular homeostasis and survival. (Right) During MASLD progression, KCs undergo excessive glycolysis enhancement, which accelerates KC death.
Author response image 1
Detection of liver fibrosis in MASLD mouse models.
Male wild-type C57BL/6J mice were fed a high-fat, high-cholesterol diet (HFHC) for 16 weeks or a high-fat diet (HFD) for 20 weeks to induce MASLD. Mice fed a normal chow diet (NCD) served as controls. (A) Sirius Red staining of liver sections was performed to assess collagen deposition and fibrosis during MASLD progression. Scale bar, 20 μm. (B) Western blot analysis of liver tissue lysates showing α-smooth muscle actin (α-SMA) expression as a marker of hepatic stellate cell activation and liver fibrosis.
Author response image 2
Assessment of KCs death on full liver tissue slice.
(A) Immunofluorescence staining was performed to detect Kupffer cell (KC) death in liver sections from mice fed an MCD diet for 6 weeks. Cell death was assessed by TUNEL staining (green), and KCs were identified by TIM4 staining (red). Nuclei were counterstained with DAPI (blue). Representative whole-tissue view is shown. Scale bars, 1mm.
Author response image 3
Chi3l1 limits glucose uptake by Kupffer cells in vivo.
(A) Measurement of 2-NBDG (a fluorescent glucose analog) uptake by KCs in vivo. WT and Chil1-/- mice, either untreated or supplemented with rChi3l1, were injected intraperitoneally with 12 mg/kg 2-NBDG. After 45mins, KCs were isolated and glucose uptake assessed by spectrophotometry. (B) Representative immunofluorescence images of liver sections stained for TIM4 (red) and 2-NBDG uptake (green) to visualize glucose uptake by KCs in situ. Scale bar = 10 µm (zoom). Quantification is shown as the percentage of TIM4+ cells that are also 2-NBDG+. Representative images were shown in B. One-way ANOVA was performed in A, B. P value is as indicated.
Tables
Appendix 1—key resources table
| Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Antibody | Rat monoclonal anti-F4/80 (APC) antibody | Invitrogen | Cat# 17-4801-82; RRID:AB_2784648 | Flow cytometry (1:100) |
| Antibody | Rat monoclonal anti-CD45 (eFluor450) antibody | Invitrogen | Cat# 48-0451-82; RRID:AB_1518806 | Flow cytometry (1:100) |
| Antibody | Rat monoclonal anti-TIM-4 (PE) antibody | Invitrogen | Cat# 12-5866-82; RRID:AB_1257163 | Flow cytometry (1:100) |
| Antibody | Rat monoclonal Anti-CD16/CD32 | Invitrogen | Cat# 14-0161-86; RRID:AB_467135 | Flow cytometry (1:100) |
| Antibody | Rat monoclonal anti-CD11b (PerCP/Cyanine5.5) antibody | BioLegend | Cat# 101228; RRID:AB_893232 | Flow cytometry (1:100) |
| Antibody | Ly-6G monoclonal antibody (1A8-Ly6g) PE-Cyanine7 | Invitrogen | Cat# 25-9668-82; RRID:AB_2811793 | Flow cytometry (1:100) |
| Antibody | Mouse monoclonal anti-Clec4f (Alexa Fluor 647) antibody | BioLegend | Cat# 156804; RRID:AB_2814082 | IF (1:300) |
| Antibody | Mouse monoclonal anti-TIM4 (Alexa Fluor 647) antibody | BioLegend | Cat# 130008; RRID:AB_2271648 | IF (1:300) |
| Antibody | Mouse monoclonal anti-β-actin antibody | Proteintech | Cat# 66009–1-lg; RRID:AB_2687938 | WB (1:1000) |
| Antibody | Cleaved caspase 3 rabbit antibody | Cell Signaling Technology | Cat# 9664S; RRID:AB_2070042 | IF (1:300), WB (1:1000) |
| Antibody | Caspase 3 rabbit antibody | Cell Signaling Technology | Cat# 9662S; RRID:AB_331439 | WB (1:1000) |
| Antibody | HNF4α rabbit antibody | Abcam | Cat# ab181604; RRID:AB_2890918 | IF (1:400) |
| Antibody | Desmin Polyclonal antibody | Proteintech | Cat#16520-1-AP | IF (1:400) |
| Antibody | IBA1 rabbit antibody | Fujifilm | Cat# 019-19741; RRID:AB_839504 | IF (1:300) |
| Antibody | Glutamine Synthetase Polyclonal antibody | Proteintech | Cat#11037-2-AP | IF (1:400) |
| Antibody | Ki67 Rabbit antibody | Abcam | Cat# ab15580; RRID:AB_443209 | IF (1:300) |
| Antibody | Rabbit polyclonal Anti-YKL-40/CHI3L1 antibody | Abcam | Cat# ab180569; RRID:AB_2891040 | IF (1:400) |
| Antibody | Peroxidase-conjugated Affinipure Goat Anti-Rabbit IgG (H+L) | Jackson | Cat# 111-035-003; RRID:AB_2313567 | WB (1:2000) |
| Antibody | Peroxidase-conjugated Affinipure Goat Anti-Mouse IgG (H+L) | Jackson | Cat# 115-035-003; RRID:AB_10015289 | WB (1:2000) |
| Antibody | Alexa Fluor 488-conjugated Affinipure Goat Anti-Mouse IgG+IgM (H+L) | Jackson | Cat# 115-545-044; RRID:AB_2338844 | IF (1:600) |
| Antibody | Alexa Fluor 568-goat anti-rabbit IgG (H+L) cross-adsorbed secondary antibody | Invitrogen | Cat# A11011; RRID:AB_143157 | IF (1:600) |
| Chemical compound, drug | FBS | VivaCell | Cat# C04001-500 | |
| Chemical compound, drug | Phosphate Buffered Saline | VivaCell | Cat# C3580-0500 | |
| Chemical compound, drug | DMEM (High glucose) | VivaCell | Cat# C3113-0500 | |
| Chemical compound, drug | DMEM (No glucose) | Sigma | Cat# D5030 | |
| Chemical compound, drug | 13C-Glucose | Cambridge Isotope Laboratories | Cat# CLM-1396-1 | |
| Chemical compound, drug | Dialysis fetal bovine serum | Abcam | Cat# DIA0500 | |
| Chemical compound, drug | Sodium pyruvate | Sangon Biotech | Cat# A501259-0100 | |
| Chemical compound, drug | Penicillin-Streptomycin Solution | VivaCell | Cat# C3421-0100 | |
| Chemical compound, drug | Cell Dissociation Solution | Sartorius | Cat# 03-079-1B | |
| Chemical compound, drug | β-Mercaptoethanol | Sigma | Cat# M3148 | |
| Chemical compound, drug | Eosin Y (water soluble) | Aladdin | Cat# E141405 | |
| Chemical compound, drug | Hematoxylin | BBI | Cat# A600701-0050 | |
| Chemical compound, drug | Oil Red O | Solarbio | Cat# IO1720 | |
| Chemical compound, drug | Sirius Red | Sangon Biotech | Cat# A500684-0500 | |
| Chemical compound, drug | High-effect paraffin-ceresin | Shanghai Hualing Rehabilitation Equipment Manufacturing Plant | Cat# N/A | |
| Chemical compound, drug | 10% Neutral Formalin Fix Solution | BBI | Cat# E672001-0500 | |
| Chemical compound, drug | Xylene | Tianjin Zhiyuan Chemical Reagents Co., Ltd. | Cat# N/A | |
| Chemical compound, drug | Neutral balsam | Solarbio | Cat# G8590 | |
| Chemical compound, drug | Isopropanol | Sangon Biotech | Cat# A507048-0500 | |
| Chemical compound, drug | Tissue-Tek OCT compound | SAKURA | Cat# REF:4583 | |
| Chemical compound, drug | Paraformaldehyde | Sangon Biotech | Cat# A500684-0500 | |
| Chemical compound, drug | Acetone | Chron Chemicals | Cat# N/A | |
| Chemical compound, drug | Sucrose | Sangon Biotech | Cat# A502792-0005 | |
| Chemical compound, drug | Triton X-100 | BBI | Cat# A600198-0500 | |
| Chemical compound, drug | Goat serum | VivaCell | Cat# C2530-0100 | |
| Chemical compound, drug | Tween 20 | BBI | Cat# A600560-0500 | |
| Chemical compound, drug | DAPI Staining Solution | Beyotime | Cat# C1006 | |
| Chemical compound, drug | Antifade Mounting Medium with DAPI | Beyotime | Cat# P0131 | |
| Chemical compound, drug | Omni-Easy One-Step PAGE Gel Fast Preparation Kit | Epizyme | Cat# PG213 | |
| Chemical compound, drug | SDS | BBI | Cat# A600485-0500 | |
| Chemical compound, drug | Glycine | BBI | Cat# A502065-0005 | |
| Chemical compound, drug | Tris | Solarbio | Cat# T8060 | |
| Chemical compound, drug | Methanol | Ghtech | Cat# N/A | |
| Chemical compound, drug | NON-Fat Powdered Milk | BBI | Cat# NON-Fat Powdered Milk | |
| Chemical compound, drug | Collagenase, Type 1 | Diamond | Cat# A004194-0001 | |
| Chemical compound, drug | Cytiva Percoll Centrifugation Media | Cytiva | Cat# 17089101 | |
| Chemical compound, drug | Heparin sodium from Porcine Intestinal | Sangon Biotech | Cat# A603251-0001 | |
| Chemical compound, drug | 1 M HEPES | Solarbio | Cat# H1095 | |
| Chemical compound, drug | OptiPrep | Serumwerk Bernburg | Cat# 1893 | |
| Chemical compound, drug | DNase I, RNase-free | Thermo | Cat# EN0521 | |
| Chemical compound, drug | CaCl2 | GHTECH | Cat#10043-52-4 | |
| Chemical compound, drug | MgSO4·7H2O | Sangon Biotech | Cat# A610329-0500 | |
| Chemical compound, drug | TRIzol reagent | Invitrogen | Cat# 15596018 | |
| Chemical compound, drug | UltraPure DNase/RNase-Free Distilled Water | Invitrogen | Cat# 10977015 | |
| Chemical compound, drug | Trichloromethane | Chron Chemicals | Cat# N/A | |
| Chemical compound, drug | PowerUp SYBR Green Master Mix | Applied Biosystems | Cat# A25742 | |
| Chemical compound, drug | DEPC水 | Biosharp | Cat# 701062 | |
| Chemical compound, drug | MgCl2 | GHTECH | Cat# N/A | |
| Chemical compound, drug | KCl | Sangon Biotech | Cat# A501159-0500 | |
| Chemical compound, drug | NaHCO3 | Sangon Biotech | Cat# A500873-0500 | |
| Chemical compound, drug | NaOH | BBI | Cat# A620617-0500 | |
| Chemical compound, drug | 2-DG | Sigma | Cat# D8375 | |
| Chemical compound, drug | PS48 | Sigma | Cat# P0022 | |
| Chemical compound, drug | Palmitic acid | Sigma | Cat# P0500 | |
| Chemical compound, drug | DMSO | Sangon Biotech | Cat# A100231-0500 | |
| Chemical compound, drug | Proteinase K Solution (20 mg/mL) | BBI | Cat# B600169-0002 | |
| Chemical compound, drug | Glycerol Gelatin aqueous slide mounting medium | Solarbio | Cat# S2150 | |
| Chemical compound, drug | XF basal medium | Agilent | Cat#103334-100 | |
| Chemical compound, drug | XF 200 mmol/L Glutamine solution | Agilent | Cat#103579-100 | |
| Chemical compound, drug | BD Pharmingen Stain Buer (FBS) | BD Biosciences | Cat# 554656 | |
| Chemical compound, drug | Draq7 | BD Biosciences | Cat# 564904 | |
| Chemical compound, drug | High-fat rodent diet with 1.25% cholesterol | Research Diet | Cat# d12108c | |
| Chemical compound, drug | Methionine and choline-deficient diet | Research Diet | Cat# A02082002BR | |
| Chemical compound, drug | Proteinase K Solution (20 mg/mL) | BBI | Cat# B600169-0002 | |
| Chemical compound, drug | Glycerol Gelatin aqueous slide mounting medium | Solarbio | Cat# S2150 | |
| Chemical compound, drug | XF basal medium | Agilent | Cat#103334-100 | |
| Chemical compound, drug | XF 200 mmol/L Glutamine solution | Agilent | Cat#103579-100 | |
| Chemical compound, drug | BD Pharmingen Stain Buer (FBS) | BD Biosciences | Cat# 554656 | |
| Chemical compound, drug | Draq7 | BD Biosciences | Cat# 564904 | |
| Chemical compound, drug | High-fat rodent diet with 1.25%cholesterol | Research Diet | Cat# d12108c | |
| Chemical compound, drug | Methionine and choline-deficient diet | Research Diet | Cat# A02082002BR | |
| Peptide, recombinant protein | Recombinant mouse Chi3l1 | SB | Cat# 50929M08H | |
| Commercial assay or kit | TMR (red) TUNEL Cell Apoptosis Detection Kit | Servicebio | Cat# G1502-100T | |
| Commercial assay or kit | Calcein/PI Cell Viability /Cytotoxicity Assay Kit | Beyotime | Cat# C2015M | |
| Commercial assay or kit | Alanine aminotransferase assay kit | Nanjing Jiancheng Bioengineering Institute | Cat# C009-2-1 | |
| Commercial assay or kit | Aspartate aminotransferase assay kit | Nanjing Jiancheng Bioengineering Institute | Cat# C010-2-1 | |
| Commercial assay or kit | Total cholesterol assay kit | Nanjing Jiancheng Bioengineering Institute | Cat# A111-1-1 | |
| Commercial assay or kit | Triglyceride assay kit | Nanjing Jiancheng Bioengineering Institute | Cat# A110-1-1 | |
| Commercial assay or kit | Seahorse XF Glycolysis Stress Test Kit | Agilent | Cat# 103020-100 | |
| Commercial assay or kit | PrimeScript II 1st Strand cDNA Synthesis Kit | TaKaRa | Cat# 6210B | |
| Commercial assay or kit | CytoTox 96Non-Radioactive Cytotoxicity Assay | Promega | Cat# G183A | |
| Software, algorithm | GraphPad Prism | GraphPad software | RRID:SCR_002798 | https://www.graphpad.com |
| Software, algorithm | FlowJo V10 | FlowJo Software | RRID:SCR_008520 | https://www.flowjo.com/ |
| Software, algorithm | ImageJ | National Institutes of Health | RRID:SCR_003070 | https://imagej.nih.gov/ij/ |
| Software, algorithm | SPSS | IBM SPSS software | RRID:SCR_002865 | https://www.ibm.com/ |
| Software, algorithm | Seahorse Wave | Agilent Technologies | RRID:SCR_024491 | https://www.agilent.com/ |
| Software, algorithm | ZEN microscope software | ZEISS | RRID:SCR_013672 | https://www.zeiss.com.cn/ |
| Cell line (Mus musculus, male) | NCTC clone 929 (L-929) | ATCC | CCL-1 | L929 was a gift from Dr. Guangxun Meng (Hainan Academy of Medical Sciences) |
| Strain, strain background (Mus musculus, male) | Chi3l1-/- | GemPharmatech Co., Ltd. | T014402 | Genetic modification: constitutive knockout |
| Strain, strain background (Mus musculus, male) | Chi3l1flox/flox | GemPharmatech Co., Ltd. | T013652 | Genetic modification: floxed allele (homozygous) |
| Strain, strain background (Mus musculus, male) | Clec4f cre | GemPharmatech Co., Ltd. | T036801 | Genetic modification: Cre recombinase transgene under Clec4f promoter |
| Other | Bone marrow-derived macrophage (BMDM) | This paper | N/A | Strain: C57BL/6J |
| Other | Kupffer cell (KC) | This paper | N/A | Strain: C57BL/6J |
Additional files
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Supplementary file 1
Metabolomics data of primary Kupffer cells from high-fat high-cholesterol diet (HFHC)-fed mice at 0, 4, and 8 weeks.
- https://cdn.elifesciences.org/articles/109206/elife-109206-supp1-v1.xlsx
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Supplementary file 2
Isotope tracing and metabolomics data for [U-¹³C]glucose.
- https://cdn.elifesciences.org/articles/109206/elife-109206-supp2-v1.xlsx
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Supplementary file 3
qPCR primers.
- https://cdn.elifesciences.org/articles/109206/elife-109206-supp3-v1.docx
-
MDAR checklist
- https://cdn.elifesciences.org/articles/109206/elife-109206-mdarchecklist1-v1.pdf
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Hyperactivated glycolysis drives spatially patterned Kupffer cell depletion in MASLD
eLife 14:RP109206.
https://doi.org/10.7554/eLife.109206.3
