Fig. 6

HDAC5 inhibition results in elevated levels of p65 acetylation. a Histogram of intracellular acetylated p65 (Ac-p65) levels of human CB CD34⁺ cells treated with vehicle, M344, or HDAC5 inhibitor LMK235. Representative histogram from three independent experiments is shown. b Mean fluorescence intensity (MFI) of intracellular acetylated p65 (Ac-p65) levels in vehicle, C646 (50 μM), M344-, M344 + C646-, LMK235-, LMK235 + C646-treated human CB CD34⁺ cells, as assessed by flow cytometry. Data pooled from two independent experiments are shown (n = 4, one-way ANOVA, ***p < 0.001). c Confocal imaging analysis of intracellular acetylated p65 (Ac-p65) levels of human CB CD34⁺ cells treated with vehicle, M344, or LMK235. Green indicates intracellular acetylated p65 staining; DAPI (blue) labels the cell nucleus. Representative images from two independent experiments are shown. Scale bar: 20 μm. d Acetylated p65 levels on CXCR4 promoter in vehicle-, M344-, or LMK235-treated human CB CD34⁺ cells, as assessed by a ChIP assay. Data pooled from two independent experiments are shown (n = 6, one-way ANOVA, ***p < 0.001). e HDAC5 IP samples blotted with an anti-acetylated p65 antibody. HDAC5 and actin in the whole cell lysate (WCL) serve as loading controls. For all panels, data are shown as mean ± SEM