Additional file 7 of FUS-induced circRHOBTB3 facilitates cell proliferation via miR-600/NACC1 mediated autophagy response in pancreatic ductal adenocarcinoma

Additional file 7: Figure S4. circRHOBTB3 promotes MiaPaca-2 cell autophagy by indirectly regulating NACC1 expression. a. The mCherr-GFP-LC3B labeled MiaPaca-2 cells were transfected circRHOBTB3 overexpression plasmid and NACC1 siRNA, and the autophagy flux was analyzed by confocal microscopy. Representative confocal microscopy images and quantitative data were shown. Scale bar, 20 μm. b. The autophagosome in MiaPaca-2 cells were detected by transmission electron microscope (TEM) upon circRHOBTB3 overexpression and NACC1 knockdown by siRNAs and corresponding negative control groups. The samples were imaged at 6000× and 15,000× magnification. The arrows indicate autophagosomes in PDAC cells. c-d. The protein levels of LC3B II, p62, phosphorylated AKT(p-AKT(T308), p-AKT(S473)), and phosphorylated mTOR(p-mTOR) in MiaPaca-2 cells with circRHOBTB3 overexpression upon NACC1 knockdown. All data are presented as the means ± SD of three independent experiments. **p < 0.01