Figure 1 .

ATG7 knockdown inhibits thrombin-induced autophagy in EC. (A) HPAEC were transfected with non-targeting siRNA (si-Con) or siRNA targeting ATG7 (si-ATG7). Cells were lysed after 48 h and analyzed by Western blot for ATG7 levels, and tubulin was used as a loading control. Error bars represent mean ± S.E. (n = 5 for each condition). (B) HPAEC transfected with si-Con or si-ATG7 as in (A) were treated with thrombin (Thr; 5 U/ml) for 1 h and lysed for Western blot analysis of LC3I/II and tubulin. Error bars represent mean ± S.E. (n = 3 for each condition). (C) si-Con and si-ATG7-transfected HPAEC were treated with thrombin (Thr; 5 U/ml) for 1 h and fixed. Cells were stained with anti-LC3 antibody (green) and DAPI (blue) to mark nuclei. Images represent three independent experiments. (D) Average LC3 puncta staining per cell represented as mean fluorescence intensity with arbitrary units (AU). Error bars represent mean ± S.E. (n = 3 with 35–45 cells were analyzed per condition).