Fig. 9

circRHOBTB3 promotes PDAC cell autophagy response via Akt/mTOR pathway. a mCherry-GFP-LC3B labeled PANC-1 were infected with circRHOBTB3 knockdown lentivirus and overexpression vector respectively, and treated with MK-2206 (5 μM,48 h) or Rapamycin (50 nM, 48 h), while circRHOBTB3 overexpression vector and SC79 (2.5 μg/ml, 48 h) or MHY1485 (5 μM, 48 h) were used for MiaPaca-2 cells. The dose of inhibitors and activators was twice as PANC-1 cells on which treated with MiaPaca-2 cells. Autophagy flux was analyzed by confocal microscopy. Representative confocal microscopy images and quantitative data were shown, with the 20 μm scale bar. b Activation or deactivation of signaling pathways were validated by detecting the protein phosphorylation level after Akt/mTOR activators or inhibitors treatment with western blot. c The protein expression levels of NACC1, LC3B I/II and p62 were detected by western blotting. All data are presented as the means ± SD of three independent experiments. **p < 0.01, ***p < 0.001