Figure 2.

HDAC6 reduces Vif and endogenous CBF-β expression. a Left, western blot of dose–response effects of HA-wt-HDAC6 and HA-HDAC6-ΔBUZ (detected with anti-HDAC6 or anti-HA abs) on over-expressed Vif and endogenous CBF-β in the absence of A3G. Lanes 1, 7 and 2, 8 are controls without or with overexpressing Vif, respectively. Right, quantitation of western blots regarding HA-HDAC6- and EGFP-HDAC6-ΔBUZ-mediated Vif degradation. b, c Cell lysates from control scrambled (SCR) or siRNA-HDAC6-treated cells, expressing recombinant or proviral Vif (pNL4.3-HIV-1). The level of expression of Vif, proviral Pr55^Gag when indicated and endogenous CBF-β are shown in each condition. Right, histogram indicates the level of expression of each protein analyzed and normalized by the amount of total α-tubulin, under any experimental condition. In all western blots, α-tubulin is the control for total protein. Acetylated α-tubulin is a read-out for functional deacetylase activity of the remaining endogenous HDAC6. Endogenous HDAC6 expression level is shown. All experiments were performed in HEK 293T cells. Data are expressed as mean ± S.E.M. of six independent experiments.