FIG. 6.

Upon PMA stimulation, the accumulation of hPKCα-D294G at the plasma membrane does not depend on the reorganization of F-actin. (A) GH3B6 cells transfected with hPKCα-D294G–GFP (top) were analyzed for F-actin localization by immunocytochemistry by using phalloidin-TRITC (bottom) in basal conditions (left) and after 1 h of PMA treatment (right). In basal conditions, F-actin was uniformly found at the plasma membrane, whereas hPKCα-D294G–GFP was cytoplasmic. Upon PMA stimulation, F-actin accumulated at the cell-cell contacts, whereas hPKCα-D294G–GFP translocated uniformly at the plasma membrane. Merged image: green, hPKCα-D294G–GFP; red, phalloidin-TRITC. Bar, 5 μm. (B) Western blot analysis, using an anti-GFP antibody, of hPKCα-D294G–GFP translocation and accumulation induced by PMA stimulation in GH3B6 cells incubated or not with phalloidin. The results show that the disorganization of the F-actin network affects neither the translocation nor the accumulation of hPKCα-D294G–GFP.