Figure 5.

Blockade of caspase-5 activation by caspase-1 inhibitors (A) and caspase-5 and -1 activation by caspase-5 knockdown (B, C). (A) hRPE cells were pretreated with caspase-1 inhibitor Z-YVAD-fmk or caspase-1 and -5 inhibitor Z-WEHD-fmk for 30 minutes and then coincubated with or without LPS (1000 ng/mL) for an additional 24 hours. Proteins from the whole hRPE cell lysates were subjected to Western blot analysis using anti–caspase-5 antibody. (B, C) Whole cell lysates from stably knockdown hTERT-RPE 1 cells with caspase-5 shRNA and control shRNA were detected by antibodies specific for pro-caspase-5 or cleaved caspase-5 or -1. Fold changes of cleaved caspase-5 and -1 were calculated by relative density between scrambled shRNA002 and caspase-5–specific shRNA3554 using densitometry after normalization to actin protein.