HIV-1 infection of human macrophages increases phosphorylation of WASP by Hck at podosomes. (A) hMDM whole-cell lysates western blot using antibodies against phosphoTyr291-WASP, WASP, and actin. (B) Quantification of the phosphoTyr291-WASP on total WASP ratio. Mean ± SD, n = 6. (C) Confocal images of hMDMs on glass. Phospho-WASP (green), p24, or Nef (red) and F-actin (blue). Arrows: accumulation of phospho-WASP at podosomes. Scale bars, 10 μm. (D) Normalized fluorescence intensity profiles of F-actin and phospho-WASP along the dotted lines in C.