Fig. 3: CMTM6 induces G0/G1 cell cycle arrest by interacting with p21 and regulating its protein levels. | Cell Death & Disease

Fig. 3: CMTM6 induces G0/G1 cell cycle arrest by interacting with p21 and regulating its protein levels.

From: CMTM6 inhibits tumor growth and reverses chemoresistance by preventing ubiquitination of p21 in hepatocellular carcinoma

Fig. 3

A Cell cycle distribution of the indicated HCC cells as assessed by flow cytometric analysis. B 293 T cells transfected with either empty vectors or vectors expressing Flag-CMTM6 were subjected to immunoprecipitation with an anti-Flag antibody, followed by silver staining. The red boxes indicated the overtly differential bands send to LC-MS/MS analysis. C Scatterplots display proteins identified by LC-MS/MS. D The best unique peptide-spectrum matches (PSM) of p21. E 293 T cells transfected with plasmids expressing either Flag-CMTM6 or Myc-p21 were subjected to immunoprecipitation with control IgG, anti-Flag (left) or anti-Myc (right) antibody. F Immunoprecipitation of HuH-7 and HepG2 cell lysates with control IgG or anti-CMTM6 antibody. G Immunoprecipitation of HuH-7 and HepG2 cell lysates with control IgG or anti-p21 antibody. H Western blot analysis of CMTM6 and p21 expression in the indicated cells. I Increasing amounts of CMTM6 plasmids were transfected into HuH-7 cells for 72 h. Protein levels of CMTM6 and p21 were then evaluated by Western blot analysis. J qPCR analysis of CMTM6 and p21 expression in the indicated cells. K Representative IHC staining images of p21 in tumor samples with high expression and low expression of CMTM6 (left). Statistical analysis of the correlation between p21 and CMTM6 expression levels detected by IHC staining (right). L Western blot analysis of p21 and CMTM6 protein expression in HCC tumor samples(left). Statistical analysis of the correlation between p21 and CMTM6 expression levels detected by Western blotting (right). *P < 0.05; **P < 0.01; ***P < 0.001; ns means non-significant.

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