FIG. 5.
Analysis of the binding and whole-cell levels of predicted TIAR target mRNAs in untreated and UVC-irradiated cells. (A) The association of endogenous TIAR with endogenous putative target mRNAs was tested using lysates prepared from RKO cells that were either left without treatment (Untr. [0 h]) or were irradiated with 25 J/m2 UVC and collected 1, 3, or 6 h afterwards. Anti-TIAR or IgG antibodies were used in IP reactions followed by the analysis of predicted target transcripts by RT-qPCR analysis of the IP material. Neg., negative control transcripts 18S rRNA and housekeeping GAPDH and UBC mRNAs; Pos., positive control MYC mRNA, a known TIAR target (32). Numbers on the right y axis indicate MYC mRNA enrichment (TIAR/IgG). (B) Total RNA was extracted from cells that were processed as described for panel A. The whole-cell levels of each mRNA were calculated and normalized to 18S rRNA levels. Data show means and standard deviations (A and B).