Figure 3

Inhibition or knockdown of PKC leads to apoptosis that is suppressed by inhibition of caspase-3. (A-E) DIC images, (F-J) fluorescent confocal images of embryos subjected to TUNEL (green) and counterstained with DAPI (blue) and (K-O) immunofluorescent confocal images of embryos stained for activated caspase-3 (green) and counterstained with DAPI (blue). (A), (F) and (K) are control embryos; (B), (G) and (L) are embryos treated with the PKC inhibitor chelerythrine from mesenchyme blastula stage; (C), (H) and (M) are embryos treated with chelerythrine and the caspase-3 inhibitor Ac-DEVD-CHO; (D), (I) and (N) are embryos injected with a SpPKC1 MASO; (E), (J) and (O) are embryos injected with the SpPKC1 MASO and treated with Ac-DEVD-CHO. The high levels of activated caspase-3 staining in panels (M) and (O) suggest that caspase-3 has been processed as a result of initiation of the apoptotic program but remains proteolytically inactive because of the presence of the inhibitor.