Figure 5.

Role of caspases and N-linked glycosylation during sphere formation and blebbishield formation in RT4P cells. (a) Role of caspases in sphere formation from blebbishields. Blebbishields were incubated with caspase inhibitors for 24 h, and the surviving spheres (inset photomicrograph) were counted. (b) Role of tunicamycin (N-linked glycosylation inhibitor) and heparin (non-specific granzyme-B inhibitor) in sphere formation from blebbishields. Blebbishields were incubated with tunicamycin or heparin for 24 h and the surviving spheres (inset photomicrograph) were counted. (c, d) Effects of caspase inhibitors, tunicamycin and heparin on blebbishield generation (c, treatments in BE medium as indicated) and subsequent sphere formation in normal MEM without inhibitors (d). Blebbishield generation in the presence of inhibitors was measured 24 h after treatment and subsequent sphere formation was measured 24 h after plating in normal MEM. During counting, blebbishields, apoptotic cells and necrotic cells were determined based on morphology shown in the inset photomicrographs. P-values in c are for blebbishields only