Fluid shear stress stimulated HDAC5 phosphorylation through a Ca2+/calmodulin–dependent pathway in endothelial cells. (A) HUVECs were exposed to flow for 0, 15, 30, 60, 120, and 240 minutes. (B-C) HUVECs were exposed to either flow or static condition for 1 hour with the pretreatment of dimethyl sulfoxide (DMSO), KN-62 (30μM), CaM inhibitory peptide (60nM), W-13 (100μM), and BAPTA/AM (30μM) + EGTA (2μM) for 30 minutes. (D) HUVECs were infected with Ad-PKD-KN or Ad-LacZ for 24 hours before the 1-hour flow or static treatment. HDAC5 phosphorylation (Ser259 and Ser498) and HDAC5 were probed by phospho-specific HDAC5 antibody and HDAC5 antibody. *P < .05 compared with control. #P < .05 compared with cells treated with flow + DMSO/Ad-LacZ. Error bars represent ±SEM.
