Figure 5.
Autophagy flux measurements suggest enhancement of macroautophagy. (A) Western blotting of LC3 in cells cultured with or without lysosomal enzyme inhibitors E64d (10 μM) and Pepstatin A (Pep.A; 10 μg/ml) in normoxic conditions. (B) Increased LC3-II levels (normalized to actin) after lysosomal enzyme inhibition (*p < 0.05; n = 4). (C) Western blotting of LC3 in cells cultured with or without lysosomal enzyme inhibitors E64d (10 μM) and Pepstatin A (10 μg/ml) in hyperoxic conditions [same groups as in (A)]. (D) Increased LC3-II levels (normalized to actin) after lysosomal enzyme inhibition (*p < 0.05; n = 4). The elevated levels of LC3-II after lysosomal enzyme inhibition indicate an increased autophagic flux.