HDAC5 interacts with p65. (A) PANC-1 cells were infected with lenti-virus expressing indicated shRNAs for 48 h. After a 48 h puromycin selection, cells were harvested for western blot analysis. (B) PANC-1 cells were transfected with indicated plasmid. 48 h after transfection, cells were harvested for co-IP assay. (C) PANC-1 cells were harvested for reciprocal co-IP of HDAC5 and p65. (D) Schematic diagram depicting a set of GST-p65 recombinant protein constructs. (E) Western blot analysis of Flag-HDAC5 proteins in PANC-1 whole cell lysate (WCL) pulled down by GST or GST-p65 recombinant proteins under the condition with or without λ-phosphatase treatment. (F) Schematic diagram depicting a set of GST-HDAC5 recombinant protein constructs. (G) Western blot analysis of p65 proteins in PANC-1 WCL pulled down by GST or GST-HDAC5 recombinant. (H-J) PANC-1 cells were infected with lenti-virus expressing indicated shRNAs for 48 h. After a 48 h puromycin selection, cells were harvested for western blot analysis (H), RT-qPCR (I) and ChIP-qPCR (J). Data are shown as mean ± SD (n = 3, n.s. not significant, ** P < 0.01, *** P < 0.001).