(A) Glucose tolerance test (GTT) assays in Tet1+/- male mice and wild-type (WT) male mice (*p < 0.05). (B) Pyruvate tolerance test (PTT) assays in Tet1+/- male mice and WT male mice (*p < 0.05). (C) …
(A) qRT-PCR analysis of the transcriptional expression of Tet1 in Tet1+/- male mice. (B) There is no change in body weight of Tet1+/- male mice and WT male mice. (C) There is no change in the food …
(A) There is no change in body weight of HFD-fed Tet1+/- male mice and HFD-fed WT male mice. (B) There is no change in the food intake between HFD-fed Tet1+/- male mice and HFD-fed WT male mice. (C) …
(A) Areas under the curve (AUC) for oral glucose tolerance test assays in Chow-fed Tet1+/- male mice and wild-type (WT) male mice. (B) Homeostatic Model Assessment for Insulin Resistance (HOMA-IR) …
(A) Transcriptional expression of selected genes in glycolysis, gluconeogenesis, and glutathione metabolism signaling pathways in TET knockdown cells using siRNA. (B and C) Representative …
(A) Respectively knockdown TET1 expression using TET1 shRNA1 and shRNA2 in LO2 cells. And the transcriptional expression of PPARGC1A, G6PC, SLC2A4, and TET1 genes were down-regulated in TET …
Supplementary Original Data for Figure 2—figure supplement 1B.
(A) Schematic representation of TET1 fragments (FL) including Flag-FL1, Flag-FL2, and Flag-FL3 (CXXC: binding CpG islands; CD: cysteine-rich domain; DSBH: double-stranded β-helix). (B) Mass spectrum …
(A) TET1 fragments were shown no interaction with HDAC1, SIRT2, and SIRT6 using protein immunoprecipitations (IPs) in TET1 fragments overexpressing cells, respectively. Each sample repeats twice.
(A) Immunofluorescence staining of the translocalization of PGC-1α or FOXO1 in EX527-treated HepG2 cells. Nuclei were stained with DAPI. (B) Western blot analysis of the fractions in EX527-treated …
(A) Comparison of the pyruvate level in liver cells between Tet1+/- mice with wild-type (WT) mice (**p < 0.01). (B) Comparison of the pyruvate level in HepG2 cells between TET1-depleted cells and …
Supplementary Original Data - Figure 5J.
(A) qRT-PCR analysis of the transcriptional expression of G6PC, PPARGC1A, and SLC2A4 were down-regulated dosage dependent on EX527. (B) qRT-PCR analysis of the transcriptional expression of G6PC, PPA…
Supplementary Original Data Figure 5—figure supplement 1C.
(A and B) qRT-PCR analysis of the transcriptional expression of the selected important genes in gluconeogenesis, including ALDH1A3 and ALDOA, with a dosage treatment of resveratrol (RSV) in HepG2 …
(A) The schedule of RSV treatment in high-fat diet (HFD)-fed Tet1+/- male mice and HFD-fed wild-type (WT) male mice. (B) The blood glucose levels in Tet1+/- male mice and WT male mice with fasting …
(A) Western blot analysis of the fractions in TET1 knockdown HepG2 cells using indicated antibodies. (B) Comparison of the effect on glycolysis among control HepG2 cells, …
(A) Western blot analysis of the fractions in EX527-treated HepG2 cells using antibodies as indicated. S1, S2, and P1 mean the proteins in the cytoplasm, nucleus soluble proteins, and chromatin …
(A) The schedule of metformin treatment in Tet1+/- male mice and wild-type (WT) male mice. (B) The glucose levels of Tet1+/- male mice and WT male mice before and after metformin treatment (*p < …
(A) There is no change in body weight of metformin-treated WT male mice compared to the control. (B) There is no change in body weight of metformin-treated Tet1+/- male mice compared to the control.
Model showing the mechanism of the AMPK-TET1-SIRT1 axis regulating the deacetylation-dependent translocalization of PGC-1α and FOXO1 from the cytoplasm to the nucleus, which contributes to glucose …
(A) IGV map presented DNA methylation signals in promoter regions of some genes, including Aldh1a3, Slc2a4, Aldoa, G6pc, Ppargc1a, and Ppargc1b. (B) The binding of 5hMC on the promoters of PPARGC1A, …
Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
---|---|---|---|---|
Gene (Homo sapiens [human]) | TET1 | GenBank | RC218608 | cDNA were purchased from Origene Corp. |
Gene (Homo sapiens [human]) | SIRT1 | GenBank | NM_012238 | |
Gene (Homo sapiens [human]) | SIN3A | GenBank | NM_001145357 | |
Cell line (Homo sapiens [human]) | HepG2 cell line | ATCC | HB-8065 | |
Cell line (Homo sapiens [human]) | LO2 cell line | Cellosaurus | CVCL_6926 | |
Cell line (Homo sapiens [human]) | HEK293T cell line | ATCC | CRL-11268 | |
Transfected construct (human) | TET1 shRNA #1; shRNA #2 | lentivirus vector pLKO_SHC201 | Cat# SHC201 | Sequence: shRNA#1: CCCAGAAGATTT AGAATTGAT shRNA#2: CCTCCAGTCTT AATAAGGTTA |
Transfected construct (human) | siRNA to TET1(SMARTpool) | Dharmacon/Thermo Fisher Scientific | Cat# D-014635 | |
Transfected construct (human) | siRNA: non-target in control (GL2 Duplex) | Dharmacon/Thermo Fisher Scientific | 1022070 | |
Sequence-based reagent | G6PC. S | This paper | RT-PCR primers | GCAGGTGTATAC TACGTGATGGT |
Sequence-based reagent | G6PC. A | This paper | RT-PCR primers | GACATTCAAGC ACCGAAATCTG |
Sequence-based reagent | PPARGC1A. S | This paper | RT-PCR primers | TGACTGGCGTC ATTCAGGAG |
Sequence-based reagent | PPARGC1A. A | This paper | RT-PCR primers | CCAGAGCAG CACACTCGAT |
Sequence-based reagent | SLC2A4. S | This paper | RT-PCR primers | GGGAAGGAAA AGGGCTATGCTG |
Sequence-based reagent | SLC2A4. A | This paper | RT-PCR primers | CAATGAGGAAC CGTCCAAGAATG |
Sequence-based reagent | TET1. S | This paper | RT-PCR primers | GGAATGGAAG CCAAGATCAA |
Sequence-based reagent | TET1. A | This paper | RT-PCR primers | ACTCCCTAAG GTTGGCAGTG |
Sequence-based reagent | PCK1.S | This paper | RT-PCR primers | ATCCCCAAAAC AGGCCTCAG |
Sequence-based reagent | PCK1. A | This paper | RT-PCR primers | ACGTACATGGT GCGACCTTT |
Sequence-based reagent | ACTB. S | This paper | RT-PCR primers | TACTGCCCTG GCTCCTAGCA |
Sequence-based reagent | ACTB. A | This paper | RT-PCR primers | GCCAGGATAGA GCCACCAATC |
Antibody | Anti-SIRT1 (Rabbit polyclonal) | Millipore | Cat# 05–1243 | WB (1:2000) |
Antibody | Anti-TET1 (Rabbit polyclonal) | Millipore | Cat# 09–872 | WB (1:1000) |
Antibody | Anti-SIN3A (Rabbit polyclonal) | Abcam | Cat# ab3479 | WB (1:2000) |
Antibody | Anti-AMPK (Rabbit polyclonal) | Abcam | Cat# ab3759 | WB (1:2000) |
Antibody | Anti-AMPK T172 (Rabbit polyclonal) | Cell Signaling | Cat# 2535 | WB (1:2000) |
Antibody | Anti-p53 (Rabbit polyclonal) | Abcam | Cat# ab26 | WB (1:2000) |
Antibody | Anti-p53 acetyl 382 (Rabbit polyclonal) | Abcam | Cat# ab75754 | WB (1:2000) |
Antibody | Anti-FOXO1 (Rabbit polyclonal) | Abcam | Cat# ab52857 | WB (1:2000)IF (1:500) |
Antibody | Anti-PGC-1α (Rabbit polyclonal) | Abcam | Cat# ab54481 | WB (1:2000)IF (1:500) |
Antibody | Anti-Histone H3 (Mouse monoclonal) | Abcam | Cat# ab1791 | WB (1:4000) |
Antibody | Anti-lysine acetylated antibody (Rabbit polyclonal) | Cell Signaling | Cat# 9441 | WB (1:2000) |
Antibody | Anti-LaminB (Rabbit polyclonal) | Santa Cruz | Cat# sc-6216 | WB (1:4000) |
Antibody | Anti-GAPDH (Mouse monoclonal) | Santa Cruz | Cat# sc-32233 | WB (1:4000) |
Antibody | Anti-5hMC (Rabbit polyclonal) | Active Motif | Cat# AB_2630381 | IP 4 µg/1 mg total lysate |
Chemical compound, drug | Metformin | Sigma Aldrich | Cat# PHR108 | |
Chemical compound, drug | AICAR | Sigma Aldrich | Cat# A9978 | |
Chemical compound, drug | RSV | Sigma Aldrich | Cat# R5010 | |
Chemical compound, drug | EX527 | Sigma Aldrich | Cat# E7034 | |
Chemical compound, drug | SRT2104 | SellectChem | Cat# S7792 | |
Commercial assay or kit | Pyruvate Assay Kit | Sigma Aldrich | Cat# MAK071 | |
Commercial assay or kit | Glucose (Go) Assay Kit | Sigma Aldrich | Cat# GAGO20 | |
Commercial assay or kit | SIRT1 Activity Assay Kit | Abcam | Cat# ab156065 | |
Software, algorithm | SPSS; Image J | SPSS; Image J | RRID:SCR_002865; RRID:SCR_003070 | |
Strain, strain background (Escherichia coli) | BL21(DE3) | Sigma Aldrich | CMC0016 | |
Other | DAPI stain | Invitrogen | D1306 | (1 µg/ml) |
Other | Lipo3000 | Invitrogen | Cat# L3000001 | |
Other | Tet1 Knockout mice | Jackson | Cat# 017358 |
List of primers.