Figure 4.

Co-immunoprecipitation of HDAC6 and A3G. a Cell lysates from control (empty pcDNA3.1 vector) or A3G-3xHA-expressing HEK 293T cells were subjected to co-immunoprecipitation (co-IP) with anti-HA Ab, followed by immunoblotting with HDAC6 and HA Abs. b Cell lysates from control [as in (a)] or A3G-3xHA-expressing cells were subjected to co-IP with anti-HDAC6 Ab, followed by immunoblotting with HDAC6 and HA Abs. c Cell-lysates from EGFP-wt-HDAC6-expressing or A3G-3xHA/EGFP-wt-HDAC6-co-expressing cells were subjected to co-IP with anti-HA Ab, followed by immunoblotting with EGFP and HA Abs. d Cell-lysates from A3G-3xHA/EGFP-wt-HDAC6- or A3G-3xHA/EGFP-HDAC6-ΔBUZ-co-expressing cells were subjected to co-IP with anti-HDAC6 Ab, followed by immunoblotting with EGFP and HA Abs. e Cell-lysates from A3G-Flag- or A3G-myc-expressing cells or cells co-expressing HA-wt-HDAC6 with each of the two A3G constructs were subjected to co-IP with anti-myc or anti-Flag Ab, followed by immunoblotting with HDAC6 or A3G Abs. f Cell-lysates from control cells (overexpressing recombinant Vif) or A3G-3xHA/Vif-co-expressing cells were subjected to co-IP with anti-HA Ab, followed by immunoblotting with Vif and HA Abs. From a to f, input expression levels of endogenous HDAC6 or overexpressed constructs are shown in cell lysates-western blots from HEK 293T cells. α-tubulin is the control for total protein. When detected, an IgG_2a heavy chain of IgG used to pull-down is indicated. Data are representative of three independent experiments.