Figure 6.

BAG3 interacts with CR and 3′UTR of CXCR4 mRNA and increases its lifetime. (a) Schematic representation of the luciferase reporter vector bearing 5′-untranslational region (UTR), coding region (CR) or 3′UTR. (b) Control or BAG3-overexpressing MCF4 cells were transfected with the indicated luciferase reporter vector and a Renilla reporter vector. Luciferase activity was measured 2 days after transfection and Renilla activity was measured to normalize luciferase activity. (c) RIP was performed using BAG3 antibody with lysates from breast cancer cells. CXCR4 mRNA enrichment was analyzed using real-time RT-PCR. (d) Biotinylated RNA segment of CXCR4 mRNA was used to pulldown lysates from MCF7 cells, and the pulldown materials were analyzed by western blotting analysis. NS, not significant; *P<0.01