Figure 4.

Cleavage of caspase 3 (A) and 8 (B) in pancreatic cancer cell lines following rfhSP-D treatment. Pancreatic cancer cell lines were analyzed for caspase 8 and 3 activation via western blot using anti-rabbit cleaved caspase 3 and 8 (1:1,000) at 4°C overnight, followed by incubation with secondary anti-rabbit IgG HRP-conjugate (1:1,000) for 1 h at room temperature. The membrane was washed with PBST (PBS + 0.05% Tween 20) three times, 10 min each between each step. The bands were developed using 3,3′-diaminobenzidine substrate kit. The cleaved caspase 3 and 8 were detected only in the rfhSP-D treated samples of all cell lines, whereas no bands appeared in the untreated cell samples. Full-length caspase 8 bands are visible around 43 kDa. (C) Anti-GAPDH was used as a loading control.