FIGURE 5 AQUAPORIN 3.

(H₂O₂ at its proliferative concentration inhibits autophagy in lung adenocarcinoma (LUAD) cells, which was neutralized by AQP3) depletion. A, Treatment with H₂O₂ alone had no effect on the LC3 I/II ratio or p62 expression. However, when Rapa treatment for 4 h was followed by H₂O₂ treatment for 2 h, the Rapa‐induced increase in autophagy was remarkably inhibited by H₂O₂. B, C, The exogenous GFP‐LC3 sequence was transfected, and autophagosome formation was monitored. H₂O₂ treatment alone had no effect on GFP‐LC3 puncta. Rapa treatment for 6 h increased the number of GFP‐LC3 puncta, and additional H₂O₂ treatment decreased the number of GFP‐LC3 puncta (**P < .01). D, Under conditions of Rapa‐induced autophagy, AQP3 depletion alone had no influence on the LC3 I/II ratio, p62 expression, or mTOR phosphorylation but counteracted the inhibitory effects of H₂O₂ on Rapa‐induced autophagy. E, F, To better assess autophagosome formation and autophagic flux, we transfected Ad‐mRFP‐GFP‐LC3 into LUAD cells in which AQP3 was depleted. Under conditions of Rapa‐induced autophagy, H₂O₂ treatment decreased the number of yellow/red puncta. AQP3 depletion alone had no effect on yellow/red puncta but counteracted the effects of H₂O₂ (**P < .01)