Figure 2.

Ethanol inhibits autophagosome formation in mouse and human pancreatic acinar cells, both in basal conditions and ex vivo alcoholic pancreatitis. A–F: acinar cells isolated from mouse or human pancreas (see experimental procedures) were preincubated for 30 min with and without bafilomycin A1 (BafA1; 20 µM) and then treated with (A and B) indicated concentrations or (C–F) 100 mM EtOH for 4 h. Where indicated, 100 pM CCK or 10 µM carbachol (CCh) was added to the cells for the last 30 min of incubation. LC3 level was measured by IB. B, D, and F: densitometric quantification of LC3-II band was performed as described in Fig. 1. Values are means ± SE. Each symbol represents individual acinar cell preparation (n = 3–7 cell preparations/condition). *P < 0.05, **P < 0.01, ***P < 0.001; one-way ANOVA with Tukey’s multiple comparison test. CCK, cholecystokinin; EtOH, ethanol; IB, immunoblot; LC3, microtubule-associated protein-1 light chain 3.