Figure 4.

Depletion of HDAC6 or inhibition of its activity impairs viral clearance. a, protocol for cellular infection with antibody-coated AdVs. Cells were seeded at the indicated density on day 1. On day 2, AdVs carrying the GFP gene were preincubated with an antibody against hexon. Cells were incubated with AdV-antibody complexes for 48 h. On day 4, the cells were harvested and infection rate was analyzed by flow cytometry. b and c, WT and HDAC6 KO MEFs were incubated with AdV-antibody complexes. d, Western blot analysis of the knockdown efficiency of siRNAs used in (e) and (f). Ctrl, control; HD6, HDAC6. The relative HDAC6 level was determined by normalizing to the corresponding α-tubulin. e–h, HeLa cells transfected with control or HDAC6-specific siRNA without (e and f) or with (g and h) tubacin or vehicle pretreatment were incubated with AdV-antibody complexes. i, Western blot analysis of the expression efficiency of the indicated FLAG-TRIM21 plasmids used in (j and k). j and k, HeLa cells transfected with HDAC6-specific siRNA and the indicated plasmids were incubated with AdV-antibody complexes. Representative plots of GFP-positive cells detected by flow cytometry (b, e, g, and j) and quantification of GFP-positive cells (c, f, h, and k) are shown (n = 6 for each dataset). Data represent the mean ± S.D. of three independent experiments. ns, not significant; *, p < 0.05, **, p < 0.01; ***, p < 0.001.