Fig. 3

Autophagy activation contributes to ferulic acid-mediated protection against lipotoxicity. a AML-12 cells were treated with different concentrations (0, 25, 50, and 100 μM) of ferulic acid (FA) for 12 h. LC3, ATG5, ATG7, Beclin1, and p62 were detected by immunoblotting. b AML-12 cells were treated with 100 μM FA for 0, 4, or 8 h. LC3 was detected by immunoblotting. c AML-12 cells were pretreated with 20 μM chloroquine (CQ, a lysosomal acidification blocker) for 1 h with or without 100 μM FA exposure for 12 h. LC3 was detected by immunoblotting. d AML-12 cells were transfected with the mRFP-GFP-LC3 plasmid before chemical exposure. The LC3 puncta were detected by confocal microscopy after the indicated treatments. e Cells were pretreated with CQ for 1 h before FA exposure. FA (100 μM) was added 2 h before palmitic acid (PA) exposure. LDH release was measured. f Caspase-3 cleavage was detected. g Nuclear morphology was detected by Hoechst staining. All values are shown as the means ± SD from three or more independent batches of cells. Bars with different superscripts are significantly different at p ˂ 0.05