fig1
Figure 1. RIP140 regulates POLK gene expression. (A) RT-qPCR analysis of Polκ and PolI mRNA in the intestinal epithelium of mice lacking the Rip140 gene (RIPKO) and in transgenic mice overexpressing RIP140 (RIPTg) as compared to their WT littermates. The results for each gene are given in arbitrary units (AU) and expressed in fold change ± S.D. relative to WT after normalization to mouse RS9 mRNA. (B) The same as in (A) in immortalized MEFs from WT or RIPKO mice. (C) HCT116 cells were stably transfected with the pEGFP-RIP140-expressing vector (HCT-RIP140) or pEGFP alone (HCT-GFP). RIP140, POLK, and POLI mRNA levels were quantified by RT-qPCR. The results are expressed relative to GFP cells after normalization to human 28S mRNA. (D) The same as in (C) in HCT116 cells transiently transfected with a RIP140 expression vector. (E) RIP140 and POLK mRNA levels were quantified by RT-qPCR in HCT116 cells transiently transfected with siCTRL or siRIP140 siRNAs as indicated. The results are expressed as fold change ± S.D. relative to siCTRL after normalization to 28S mRNA (n = 3 independent experiments for each condition). (F) The same as in (E) performed in RKO CRC cells. (G) POLK expression analysis by Western blot of whole cell extract from RKO cells 48 h after transient siRNA transfection. Quantifications are expressed in AU after normalization to β-actin, used as a control of protein migration. A Mann-Whitney test was used for statistical analysis (ns = not significant, **P < 0.01, ***
