Journal: Advanced Science

Article Title: Lactylation‐Driven YTHDC1 Alleviates MASLD by Suppressing PTPN22‐Mediated Dephosphorylation of NLRP3

doi: 10.1002/advs.202510192

Figure Lengend Snippet: YTHDC1 is downregulated in MASLD. A) Representative immunohistochemical and H&E staining of liver tissue of patients with MASLD and normal donor liver tissue. Scale bars=100 µm. n = 16–21. B) Representative statistics of immunohistochemical staining of liver tissue and normal donor liver tissue in MASLD patients. n = 16–21. C) Analysis of the correlations between YTHDC1 level and lipid droplet‐positive areas. n = 16–21. D) Heatmap for correlation analysis of YTHDC1 and TC, TG, ALT, AST, GGT levels in plasma of healthy controls and MASLD patients. n = 16–21. E) Western blot analysis of YTHDC1 protein levels in MASLD patients and normal individuals. n = 6. F) Western blot analysis of YTHDC1 protein levels in HFD‐induced mice and golden hamster groups and at 24 h after OA induction in HepG2 and Thle‐2 cells. n = 6. G) Immunofluorescence and BODIPY representative images of YTHDC1 expression in oleic acid‐induced HepG2 and Thle‐2. Scale bars=100 µm. n = 4. Data are presented as mean ± SEM.

Article Snippet: HepG2, L02, and THLE‐2 cell lines were purchased from the following sources: HepG2 from Procell Life Science & Technology Co., Ltd.; L02 from the Type Culture Collection of the Chinese Academy of Sciences, Shanghai, China; and THLE‐2 from iCell Bioscience Inc.

Techniques: Immunohistochemical staining, Staining, Clinical Proteomics, Western Blot, Immunofluorescence, Expressing

Journal: Nature communications

Article Title: O-GlcNAcylated p53 in the liver modulates hepatic glucose production.

doi: 10.1038/s41467-021-25390-0

Figure Lengend Snippet: Fig. 8 Hepatic p53 affects postprandial hyperglycemia and is elevated in models of insulin resistance. a Hepatic OGT and p53 protein levels in WT mice fed with standard diet or high-fat diet (HFD) 60% for 4 days (n = 7). Hepatic OGT protein levels in (b) control mice and p53 LKO mice fed with HFD 60% for 4 days (n = 6) and c FLOXp53 mice injected with AAV8 expressing either GFP (n = 7) or Cre (n = 5) and fed with HFD 60% for 4 days. Blood glucose levels in Alfp-Cre± mice injected with AAV-DIO expressing either GFP or p53 refed with d chow diet (n = 11 and 12) and e HFD after overnight fasting (n = 11 and 12). Blood glucose levels in FLOXp53 mice injected with AAV8 expressing either GFP or Cre refed with f chow (n = 6 and 8) diet and g HFD after overnight fasting (n = 5 and 6). Blood glucose levels in control and p53 LKO (n = 6) mice refed with h chow (n = 5 and 7) diet and i HFD (n = 7) after overnight fasting. j Hepatic p53 and PCK1 protein levels in WT (n = 4) and db/db mice (n = 9). The area under the curve (AUC) is provided. Expression of GAPDH served as a loading control and control values were normalized to 100%. Dividing lines indicate splicings within the same gel. Data were presented as mean ± standard error mean (SEM). * denotes P < 0.05, ** denotes P < 0.01, and *** denotes P < 0.001, determined by two-tailed Student’s t-test. “n” denotes independent animals. Source data are provided as a Source Data file.

Article Snippet: To upregulate p53 protein levels in THLE-2 cells, cells were transfected with pCMV-Neo-Bam and pCMV-Neo-Bam p53 wt (Addgene plasmid #16440 and Fig. 7 Overexpression of hepatic p53 worsens insulin sensitivity. a p53 levels in THLE-2 cells maintained in complete medium (CM) (n= 5), KHH (n= 4), or KHH+ insulin (n= 4). b Protein levels of p53 and PCK1 in THLE-2 cells maintained in CM, KHH, or KHH+ insulin (n= 5). c Protein levels of p53 and PCK1 in THLE-2 cells transfected with empty plasmid or plasmid encoding p53 and then treated with insulin (n= 6). d PCK1 activity (n= 6) and e glucose production (p0 n= 4; p0+ insulin 10 nM n= 5; and pp53+ insulin 10 nM n= 6) in THLE-2 cells transfected with empty plasmid or plasmid encoding p53 cells in the absence or presence of insulin. f Protein levels of p53 and PCK1 in THLE-2 cells are maintained in KHH in the presence or absence of insulin and PUGNAc (n= 6). g Protein levels of pAKT, p53, and PCK1 in THLE-2 cells maintained in CM (n= 3) or KHH in the absence (n= 3) or presence of SC79 (n= 4). h PCK1 levels in THLE-2 cells transfected with empty plasmid or plasmid encoding p53 and then treated with SC79 (n= 6). i Protein levels of p53 and PCK1 in mice injected with empty adenovirus or p53 adenovirus (Adp53) (n= 3 and 6 per group). j Protein levels of pAKT, p53, and PCK1 in control mice or mice over-expressing p53 and then treated with saline or insulin into the cava vein (n= 6 and 7). k Blood glucose levels after IP insulin treatment (n= 8 and 10) and l protein levels of PCK1 in Alfp-Cre± mice injected with AAV8-DIO expressing either GFP or p53 and then treated with saline or insulin into the cava vein (n= 4 and 5).

Techniques: Control, Injection, Expressing, Two Tailed Test

Journal: Nature communications

Article Title: O-GlcNAcylated p53 in the liver modulates hepatic glucose production.

doi: 10.1038/s41467-021-25390-0

Figure Lengend Snippet: Fig. 9 p53 protein levels are increased in the liver of patients with T2D. Obese patients were further subclassified according to their normoglycemia (NG) or type 2 diabetes (T2D) (n = 30 human patients per group). a Hepatic p53 mRNA levels in NG or T2D patients. b Hepatic p53 and PCK1 protein levels in patients with NG and T2D. c Correlation between p53 protein levels and OGTT. d Expression of OGT, OGA, GFAT1, and GFAT2 in patients with NG and T2D. e Correlation between GFAT1 and GFAT2 expression and OGTT. Expression of GAPDH (western blot) and HPRT (qRT-PCR) served as a loading control, and control values were normalized to 100%. Dividing lines indicate splicings within the same gel. Data were presented as mean ± standard error mean (SEM). * denotes P < 0.05, ** denotes P < 0.01, and *** denotes P < 0.05 determined by two-tailed Student’s t-test. Source data are provided as a Source Data file.

Article Snippet: To upregulate p53 protein levels in THLE-2 cells, cells were transfected with pCMV-Neo-Bam and pCMV-Neo-Bam p53 wt (Addgene plasmid #16440 and Fig. 7 Overexpression of hepatic p53 worsens insulin sensitivity. a p53 levels in THLE-2 cells maintained in complete medium (CM) (n= 5), KHH (n= 4), or KHH+ insulin (n= 4). b Protein levels of p53 and PCK1 in THLE-2 cells maintained in CM, KHH, or KHH+ insulin (n= 5). c Protein levels of p53 and PCK1 in THLE-2 cells transfected with empty plasmid or plasmid encoding p53 and then treated with insulin (n= 6). d PCK1 activity (n= 6) and e glucose production (p0 n= 4; p0+ insulin 10 nM n= 5; and pp53+ insulin 10 nM n= 6) in THLE-2 cells transfected with empty plasmid or plasmid encoding p53 cells in the absence or presence of insulin. f Protein levels of p53 and PCK1 in THLE-2 cells are maintained in KHH in the presence or absence of insulin and PUGNAc (n= 6). g Protein levels of pAKT, p53, and PCK1 in THLE-2 cells maintained in CM (n= 3) or KHH in the absence (n= 3) or presence of SC79 (n= 4). h PCK1 levels in THLE-2 cells transfected with empty plasmid or plasmid encoding p53 and then treated with SC79 (n= 6). i Protein levels of p53 and PCK1 in mice injected with empty adenovirus or p53 adenovirus (Adp53) (n= 3 and 6 per group). j Protein levels of pAKT, p53, and PCK1 in control mice or mice over-expressing p53 and then treated with saline or insulin into the cava vein (n= 6 and 7). k Blood glucose levels after IP insulin treatment (n= 8 and 10) and l protein levels of PCK1 in Alfp-Cre± mice injected with AAV8-DIO expressing either GFP or p53 and then treated with saline or insulin into the cava vein (n= 4 and 5).

Techniques: Expressing, Western Blot, Quantitative RT-PCR, Control, Two Tailed Test

Journal: Molecular Oncology

Article Title: Benzylidene‐indolinones are effective as multi‐targeted kinase inhibitor therapeutics against hepatocellular carcinoma

doi: 10.1016/j.molonc.2014.04.008

Figure Lengend Snippet: Anti‐proliferative effects of 47 on HCC cell lines (A) HuH7, (B) HepG2 and (C) THLE2 cells on 96‐well format were treated with 0‐10 µM (■), Sunitinib (●) or Sorafenib (▲) for 72 h. Viability was ...

Article Snippet: HepG2, Hep3B and PLC/PRF/5 were maintained in MEM, THLE2 in BEGM (Lonza, Basel, Switzerland), and all other cell lines in DMEM.

Techniques: