Fig. 6: GATA4 facilitates HDAC1-mediated deacetylation of p65 by enhancing the interaction between p65 and HDAC1.

A, B HEK293T cells were transfected with indicated plasmids, followed by immunoprecipitation with anti-Flag affinity magnetic beads and western blot with anti-AcK Ab. C HEK293T cells were transfected with indicated plasmids, and incubated with or without TNFα (5 μM for 1 h), followed by immunoprecipitation with anti-Flag affinity magnetic beads and western blot with anti-AcK Ab. D The luciferase reporter assay showed the effect of HDAC1 in HEK293T cells on MMP9 promoter with or without GATA4. Data are the means ± SDs from three determinations. *p < 0.05. E Lysates in T47D cells were immunoprecitated using anti-GATA4 Ab. Purified cell extracts were tested by western blot with anti-HDAC1 Ab. F HEK293T cells were co-transfected with HA-HDAC1 and Flag-GATA4, and lysates were immunoprecitated using anti-Flag affinity magnetic beads. Purified cell extracts were tested by western blot with anti-HA Ab. G GST-pulldown assay showed the purified GST-GATA4 in E. coli BL21 had physical interaction with HA-HDAC1 in HEK293T cells. H T47D cells were stained with an anti-GATA4 Ab (green) and an anti-HDAC1 Ab (red). Nuclei were stained with DAPI (blue), followed by visualization with confocal microscopy. I HEK293T cells were transfected with indicated plasmids, followed by immunoprecipitation with anti-Flag affinity magnetic beads and western blot with anti-AcK Ab. J HEK293T cells were co-transfected with HA-HDAC1, GFP-p65, and Flag-GATA4, and lysates were immunoprecitated using anti-GFP Ab. Purified cell extracts were tested by western blot with anti-HA and anti-Flag Ab.