Hepatic NF-kB-inducing kinase (NIK) suppresses mouse liver regeneration in acute and chronic liver diseases
- University of Michigan Medical School, United States
- University of Campinas, Brazil
- China Pharmaceutical University, China
- Wuhan University, China
Figures
Figure 1
Hepatocyte-specific ablation of NIK accelerates reparative hepatocyte proliferation.
NIKf/f (n = 7) and NIKΔhep (n = 7) male mice (8 weeks) were subjected to PHx, and livers were harvested 48 hr or 96 hr later. (A) Representative immunostaining of liver sections (48 hr after PHx) …
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Figure 1—source data 1
Hepatic NIK deficiency accelerates liver regeneration.
- https://doi.org/10.7554/eLife.34152.003
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Figure 1—source data 2
PHx increases hepatocyte replications.
- https://doi.org/10.7554/eLife.34152.004
Figure 2 with 1 supplement
Hepatic NIK deficiency upregulates cyclin D1 without altering NF-kB1, Akt, and MAPK pathways in the liver.
NIKf/f and NIKΔhep male mice (8 weeks) were subjected to PHx. (A–B) Liver extracts were immunoblotted with anti-cyclin D1 antibody (48 hr after PHx). Cyclin D1 levels were quantified and normalized …
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Figure 2—source data 1
Hepatic NIK regulates hepatocyte cell cycle progression.
- https://doi.org/10.7554/eLife.34152.008
Figure 2—figure supplement 1
Effect of PHx on liver NIK pathway activation.
PHx was performed on C57BL/6 male mice. (A) Liver NIK mRNA abundance was measured by qPCR (normalized to 36B4 levels, n = 3 per group). (B–C) Liver extracts were immunoblotted with antibodies …
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Figure 2—figure supplement 1—source data 1
PHx stimulates hepatic NIK expression.
- https://doi.org/10.7554/eLife.34152.007
Figure 3
NIK inhibits the JAK2/STAT3 pathway.
(A) Liver extracts were prepared from NIKf/f and NIKΔhep males 4 hr after PHx and immunoblotted with anti-phospho-JAK2 and anti-phospho-STAT3 antibodies. Phosphorylation of JAK2 (pTyr1007/1008) and …
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Figure 3—source data 1
NIK inhibits the JAK2/STAT3 pathway.
- https://doi.org/10.7554/eLife.34152.010
Figure 4
Ablation of hepatocyte IKKα accelerates hepatocyte reparative proliferation.
(A) Tissue extracts were immunoblotted with anti-IKKα or anti-α-tubulin antibodies. (B–F) IKKαf/f (n = 6) and IKKαΔhep (n = 6) male littermates were subjected to PHx, and livers were harvested 48 hr …
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Figure 4—source data 1
Hepatic IKKα regulates liver regeneration.
- https://doi.org/10.7554/eLife.34152.012
Figure 5 with 1 supplement
IKKα inhibits the JAK2/STAT3 pathway.
(A–B) Liver extracts were prepared 4 hr after PHx and immunoblotted with anti-phospho-JAK2 and anti-phospho-STAT3 antibodies. Phosphorylation of JAK2 (pTyr1007/1008) and STAT3 (pTyr705) was …
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Figure 5—source data 1
IKKα regulates the JAK2/STAT3 pathway.
- https://doi.org/10.7554/eLife.34152.017
Figure 5—figure supplement 1
The effect of PHx on activation of liver IKKα and JAK2/STAT3 pathways.
IKKαf/f and IKKαΔhep male mice were subjected to PHx, and livers were harvested on days 0–7. (A) Liver extracts were immunoblotted with the indicated antibodies. (B) Phosphorylation of JAK2 …
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Figure 5—figure supplement 1—source data 1
PHx regulates the hepatic JAK2/STAT3 pathway.
- https://doi.org/10.7554/eLife.34152.015
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Figure 5—figure supplement 1—source data 2
Regulation of the JAK2/STAT3 pathway by NIK/IKKα pathways.
- https://doi.org/10.7554/eLife.34152.016
Figure 6 with 1 supplement
Ablation of hepatocyte NIK reverses AAF-induced impairment in hepatocyte reparative proliferation.
(A) C57BL/6 males (8 weeks) were treated with PBS or AAF (10 mg/kg body weight, gavage) daily for 10 days. NF-kB2 p52 in liver extracts was immunoblotted with anti-NF-kB2 antibody (normalized to …
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Figure 6—source data 1
Hepatic NIK regulates hepatocyte proliferation in AAF-treated mice.
- https://doi.org/10.7554/eLife.34152.022
Figure 6—figure supplement 1
Hepatic NIK inhibits reparative hepatocyte proliferation.
(A) NIKf/f and NIKΔhep males were treated with PBS or AAF (10 mg/kg body weight) for 10 days and then subjected to PHx. Resected liver weight and liver weight 2 days post-PHx were normalized to body …
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Figure 6—figure supplement 1—source data 1
Baseline hepatocyte proliferation in AAF-treated mice.
- https://doi.org/10.7554/eLife.34152.020
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Figure 6—figure supplement 1—source data 2
Baseline hepatocyte proliferation in HFD-fed mice.
- https://doi.org/10.7554/eLife.34152.021
Figure 7
Hepatic NIK deficiency corrects impaired hepatocyte reparative proliferation in mice with NAFLD.
(A–B) C57BL/6 males (8 weeks) were fed a normal chow diet (n = 5) or a HFD (n = 5) for 10 weeks. (A) Liver TAG levels (normalized to liver weight). (B) NF-kB2 p52 in liver extracts was immunoblotted …
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Figure 7—source data 1
Hepatic NIK regulates hepatocyte proliferation in HFD-fed mice.
- https://doi.org/10.7554/eLife.34152.024
Tables
Key resources table
| Reagent type | Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Antibody | Ki67 | Vector lab | VP-RM04 | 1:100 |
| Antibody | NIK | Abcam | ab191591 | 1:2000 |
| Antibody | IKK beta | Cell Signaling Technology | 8943 | 1:5000 |
| Antibody | IKK alpha | Cell Signaling Technology | 2682 | 1:5000 |
| Antibody | p-IKKa/b | Cell Signaling Technology | 2697 | 1:5000 |
| Antibody | STAT3 | Santa Cruz | sc-8019 | 1:1000 |
| Antibody | p-STAT3 | Cell Signaling Technology | 9145 | 1:5000 |
| Antibody | JAK2 | Cell Signaling Technology | 3230 | 1:5000 |
| Antibody | p-JAK2 1007/1008 | Cell Signaling Technology | 3776 | 1:5000 |
| Antibody | Myc | Santa Cruz | sc-40 | 1:1000 |
| Antibody | Flag | Sigma | F1804 | 1:5000 |
| Antibody | p85 | Home-raised | N/A | 1:5000 |
| Antibody | α-tubulin | Santa Cruz | sc-5286 | 1:1000 |
| Antibody | JNK | Cell Signaling Technology | 9258 | 1:5000 |
| Antibody | p-JNK | Cell Signaling Technology | 4668 | 1:5000 |
| Antibody | ERK1/2 | Cell Signaling Technology | 9102 | 1:5000 |
| Antibody | p-ERK1/2 | Cell Signaling Technology | 4370 | 1:5000 |
| Antibody | NF-kB2 | Cell Signaling Technology | 4882 | 1:5000 |
| Antibody | p65 | Cell Signaling Technology | 8242 | 1:5000 |
| Antibody | p-p65 | Cell Signaling Technology | 3033 | 1:5000 |
| Antibody | IkB alpha | Cell Signaling Technology | 4812 | 1:5000 |
| Antibody | p-IkB alpha | Cell Signaling Technology | 9246 | 1:5000 |
| Antibody | AKT | Cell Signaling Technology | 4091 | 1:5000 |
| Antibody | p-Akt | Cell Signaling Technology | 4060 | 1:5000 |
| Antibody | Cyclin D1 | Cell Signaling Technology | 2978 | 1:5000 |
| Antibody | F4/80 | eBioscience | 14–4801 | 1:100 |
| Antibody | HNF4 alpha | Santa Cruz | sc-8987 | 1:100 |
| Antibody | CK8 | Developmental Studies Hybridoma Bank | Troma I | 1:100 |
| Antibody | BrDU | Cell Signaling Technology | 5292 | 1:100 |
Additional files
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Transparent reporting form
- https://doi.org/10.7554/eLife.34152.025
