Figure 6

Both PKCα and PKCδ are required for αvβ3-mediated invasion. A) C8161.9 cells were transfected with scrambled (Scram), PKCδ siRNA (si1δ), or two PKCα siRNAs (si2α, si3α) and siRNA-transfected cells were plated on vitronectin for 2 hours and the extent of stress fiber and focal adhesion formation was monitored by staining with phalloidin (red) and immunostaining with anti-paxillin (green) respectively. Nuclei were counterstained with Hoescht (blue). White arrows indicate actin stress fibers around the perinuclear area in PKCα siRNA-treated cells. B) C8161.9 cells were transfected with scrambled (Sc), PKCδ (i1δ), si2α PKCα (i2α), or si3α PKCα (i3α) siRNAs. The levels of PKCα, PKCδ, and PKCε in siRNA transfected cells were measured by immunoblotting with isoform-specific PKC antibodies. Total levels of tubulin were also monitored (tubulin). C) C8161.9 cells were transfected with scrambled (Sc), PKCα siRNAs si3α (si3a), si2α (si2a), or PKCδ siRNA (si1d) and tested for their ability to invade VN-enriched Matrigel. Data represents at least 4 independent experiments. D) M14 cells were infected with viruses expressing nothing (V), PKCα (α), PKCδ (δ), Src, or PKCα and PKCδ (α + δ). Total levels of each protein were monitored by immunoblotting. E) Ability of the over expressing M14 cells to invade VN-enriched Matrigel was monitored. Data represents 3 independent experiments.