FIG 9.

Tumor suppressor activity of CUGBP1 is associated with repression of multiple pathways of liver cancer for the levels of splicing and expression of mRNAs. (A) Analysis of RNA-Seq data shows lists of altered gene pathways regarding the levels of splice variants and mRNA in livers of DEN-treated CUGBP1 S302A mice versus livers of DEN-treated WT mice. Pathways in red are associated with liver cancer. (B) Confirmation of changes of levels of mRNAs by qRT-PCR. (C) Western blot of c-jun protein. The filter was first probed with antibodies to c-jun and then reprobed with antibodies to CUGBP1 and β-actin. (D) Western blotting with antibodies to ph-S780-Rb. Nuclear extracts of untreated and DEN-treated (38 weeks) mice were used. The three lower images show results of co-IP studies. E2F1 was immunoprecipitated and Rb and E2F1 were examined in these IPs. The image labeled Rb dark shows a dark exposure of Rb membrane to detect Rb in IPs from DEN-treated mice. The bottom image shows IgG signals on the same filter.