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. 2020 Jun 8;117(25):14376–14385. doi: 10.1073/pnas.1921618117

Fig. 1.

Fig. 1.

LC3 and LC3-II are induced by 15LO1-dependent mechanisms in HAECs in vitro under IL-13 stimulation. (A) IL-13 induced LC3 mRNA expression in a dose-dependent manner. HAECs under ALI were stimulated with IL-13 1 and 10 ng/mL for 7 d. LC3 mRNA was analyzed by RT-PCR with beta-glucuronidase (GUSB) as the internal control (*as compared to no-IL-13 control). (B) Representative western blot showing IL-13–induced LC3-II protein expression in HAECs in vitro. HAECs under ALI were stimulated with IL-13 for 7 d. Total cell lysate protein was collected for western blot analysis. (C) Representative western blot showing HCQ further increased IL-13–induced LC3-II. HAECs were all stimulated with IL-13 for 7 d before treatment with or without HCQ (10 μM, overnight). (D) 15LO1 siRNA (siALOX15) transfection suppressed IL-13–induced LC3-II protein expression. HAECs transfected with siALOX15 were all stimulated with IL-13 for 5 d, with scramble siRNA as the negative control. (E) The 15LO1 inhibitor BLX2477 (BLX) suppresses IL-13–induced LC3-II. HAECs were all stimulated with IL-13 for 7 d before treatment with BLX2477 (2 μM, overnight), with DMSO as the control. GAPDH, glyceraldehyde-3-phosphate dehydrogenase.