Fig. 2.

NFV reduces active caspase 3 staining and cytochrome c (Cyto.c) leakage into the cytosol during pancreatitis. Pancreas was harvested 1 h after the last of a series of 12 hourly intraperitoneal injections of saline (A), 50 μg/kg caerulein (B), animals pretreated with water before administration of caerulein (C), and animals pretreated with NFV/ritonovir (RTV) (N/R; D) or Z-VAD-fmk (ZVAD; E) before administration of caerulein. These were fixed and stained for active caspase-3 and accessed as described in materials and methods. Representative ×400 images are shown. F: % active caspase-3 positive cells expressed as means ± SE. Immunoblots of cytoplasmic fractions (G) and mitochondrial fractions (H) blotted for cytochrome c (top) and then stripped and blotted for HSP70 (G, middle and H, bottom) which was used a control for lane loading of the mitochondrial fractions. Since there was no leakage of heat shock protein (HSP)70 into the cytosolic fractions, PCNA was used as a control for lane loading of the cytosolic fractions as described in materials and methods (G, bottom). Each column represents pancreatic protein amounts in the respective fractions of pancreatic tissue after the different treatments. The 4 different treatments were control animals, 3-h caerulein pancreatitis, 3-h pancreatitis prophylaxed with vehicle, and 3-h pancreatitis prophylaxed with NFV/RTV.