FIGURE 4.

Inhibition of autophagy rescued endothelial cells from PA-induced death. A, representative Western blot showing LC3 cleavage of cells that were incubated for 8 h with BSA alone, 0.5 mm OA, or 0.5 mm PA in the absence (−) and presence (+) of 10 μm wortmannin. B, statistical data of LC3 cleavage from Western blots shown in panel f (n = 3 for all conditions). *, p < 0.05 versus control. C, representative Western blot showing LC3 cleavage of cells that were incubated for 8 h with BSA alone or 0.5 mm PA in the absence (−) and presence (+) of 10 mm 3-MA. D, statistical data of LC3 cleavage from Western blots shown in panel f (n = 3 for all conditions). *, p < 0.05 versus control. E, columns represent average cell viabilities that were determined using the MTT assay of cells not pretreated with wortmannin (left pair of columns, −Wortmannin) that were incubated for 24 h with BSA alone (left white column, n = 3) or with 0.5 mm PA complexed to BSA (left black column, n = 3), and cells pretreated with 10 μm wortmannin (right pair of columns, +Wortmannin) for 20 min prior to an incubation with either BSA alone (right white column, n = 3) or with 0.5 mm PA complexed to BSA (right black column, n = 3). *, p < 0.05 versus BSA, and #, p < 0.05 versus PA without wortmannin. F, cells were treated with 10 mm 3-MA, another specific inhibitor of PI3K III and autophagy, and cell death was analyzed by the MTT assay (n = 3 for all conditions). *, p < 0.05 versus BSA, and #, p < 0.05 versus PA without 3-MA.