FIGURE 3.

Wild type PKCα but not PKCα lacking the last three amino acids rescues wound healing in PKCα-depleted H1703 cells. Upper panel, a Western blot shows re-expression of bovine wild type (wt) HA-tagged PKCα or PKCα lacking the last three amino acids (ΔPDZ) in H1703 cells stably expressing NT or PKCα shRNA in a wound healing assay as described below. After completion of the scratch assay, cells were lysed, and lysates were probed for PKCα, HA, and β-actin. Lower panel, shown is the effect of PKCα rescue on wound healing. The indicated H1703 cells were transiently transfected with enhanced GFP (eGFP; as a marker for transfection) with or without HA-PKCα or HA-PKCα-ΔPDZ. Cells were plated at equal densities and then pretreated with mitomycin C (to inhibit proliferation) and scratched with a pipette tip, and their migration was monitored over 8–16 h. The area covered by migrating cells was quantified using ImageJ, and data points represent the mean ± S.E. of four experiments. *, significantly different from the NT shRNA + eGFP condition, p < 0.05.