Fig. 4.

DHHC3 knockdown affects endogenous β4 and α6 palmitoylation and expression. a MDA-MB-231 cells and b PC3 cells, each with mock, control and DHHC3 stable knockdown, were labeled with [³H]palmitate for 2 h. Cells were lysed in 1% Brij96 buffer and then endogenous β4 and α6 integrins were immunoprecipitated and detected from the [³H] radioactive signal (top panel) and by protein immunoblotting. In part a, the top right panel shows many unidentified palmitoylated proteins in the whole cell lysate, and the bottom right panel shows DHHC3 protein expression. c Vector and DHHC3 knockdown PC3 cells were permeabilized and stained with β4 and α6 antibodies, and visualized by confocal microscopy. V Vector control knockdown; D3 DHHC3 knockdown. Bar,20 μm. d Staining intensity (units from ImageJ program) is quantitated for two representative cells from each panel in part c