Fig. 4. BAG3 knockdown destabilizes IL-8 mRNA via miRISC complex.

a RIP was performed using Ago2 antibody, enrichment of IL-8 mRNA was analyzed using qRT-PCR. b Ago2 protein expression was analyzed using western blot. c BxPC3 cells were infected with lentivirus containing Ago2 (shAgo2), knockdown efficiency wax confirmed by western blot analysis. d–e BxPC3 cells were infected with lentivirus containing shAgo2, IL-8 mRNA was analyzed using qRT-PCR. f–g The indicated BxPC3 cells were transfected with luciferase containing IL-8 3′-UTR, luciferase activity was analyzed. h–i Control or BAG3 KD BxPC3 cells were transfected with Ago2, Ago2 expression was confirmed by western blot (h), IL-8 mRNA was analyzed using qRT-PCR i. j BxPC3 cells were co-transfected with Ago2 and luciferase construct containing IL-8 3′-UTR, luciferase activity was analyzed. N.S., not significant; *, P < 0.01