Fig. 4

SLCO4A1-AS1 interacts with Hsp90 and Cdk2 in CRC. a and b Subcellular localization of SLCO4A1-AS1 was determined by qRT-PCR (a) and FISH (b) in HCT116 cells. c Proteins retrieved from the SLCO4A1-AS1 pull-down assay were analyzed by SDS-PAGE in HCT116 cells. d Western blotting analyses of Hsp90 and Cdk2 using the proteins retrieved from the SLCO4A1-AS1 pull-down assay in HCT116 cells. e RIP assays using an anti-Cdk2 or anti-Hsp90 antibody showed that both Cdk2 and Hsp90 interact with SLCO4A1-AS1 in HCT116 cells. The qRT-PCR results of RIP assays are shown in the upper panel, and the agarose electrophoresis results of the PCR products are shown in the lower panel. f Western blotting of Cdk2 and Hsp90 in protein samples pulled down by the full-length (F2:1–1440 bp) or truncated SLCO4A1-AS1 (F3: 600–1440 bp, F4: 900–1440 bp, F5: 1–900 bp) (the upper panel), and truncation mutants of SLCO4A1-AS1 shown by agarose electrophoresis (the lower panel). g Graphic illustration of the secondary structure of SLCO4A1-AS1 predicted by ViennaRNA (http://rna.tbi.univie.ac.at/) and its corresponding regions accounting for the binding with Cdk2 and Hsp90. *P < 0.05; **P < 0.01