Fig. 8

circRHOBTB3 promotes PANC-1 cell proliferation by activating autophagy response. a The protein level of p62 and LC3B were determined by western blotting in PANC-1 cells transfected circRHOBTB3 overexpressing vector with or without 3-Methyladenine (3-MA, 2.5 mM, 24 h) treatment. b The mCherry-GFP-LC3B labeled PANC-1 cells were transfected with circRHOBTB3 overexpression vector with or without 3-MA (2.5 mM, 24 h) treatment, the LC3B puncta were analyzed by confocal microscopy. Representative images and quantitative data were shown, with 20 μm scale bar. c The transmission electron microscopy revealed the autophagosome with the magnification of 6000X and 15,000X, Arrows indicate autophagosomes in cells. Scale bar 2 μm and 1 μm respectively. d The colony formation revealed that 3-MA (2.5 mM, 24 h) treatment inhibited the increased proliferation after circRHOBTB3 overexpression. e The DNA synthesis ability was restrained by 3-MA (2.5 mM, 24 h) treatment in cells with overexpressed circRHOBTB3 as suggested by EdU incorporation assays. f The growth curve of PANC-1 cells showed reversed proliferation capability upon 3-MA (2.5 mM, 24 h) treatment in cells with circRHOBTB3 overexpression. All data are presented as the means ± SD of three independent experiments. **p < 0.01, ***p < 0.001