Fig 2: Radiosensitization and autophagy inhibition induced by GABARAP knockdown in HCC cell lines. (a) GABARAP mRNA expression levels in HLF and HuH6 cells transduced with shNT or GABARAP-targeting shRNAs (shGa and shGb) for 48 h. * p < 0.05 vs. shNT; Dunnett’s test (n = 3). (b) LC3 and GABARAP protein expression in HLF and HuH6 cells transduced with shNT or GABARAP-targeting shRNAs (shGBRPa and shGBRPb) for 48 h. (c) Clonogenicity of HLF and HuH6 overexpressing NEAT1v1 after X-ray irradiation. Following irradiation, the cells were immediately transduced with shNT, shGa, or shGb. * p < 0.05 vs. shNT; Dunnett’s test (n = 4).

Fig 3: C-2-induced autophagy is associated with JNK pathway by alleviating the suppression of Bcl-2 and Bcl-xL on Beclin-1 in bladder cancer cells. a. The phosphorylation of JNK and c-Jun were analyzed by western blotting at indicated concentration or treated with 4 µM of C-2 at indicated time points in BIU87 and EJ cells. b BIU87 and EJ cells were pretreated with 10 µM of SP600125 for 1 h and then treated with 4 µM of C-2 for an additional 24 h. The indicated proteins were detected by western blotting after immunoprecipitation with an antibody for Beclin-1. c The effects of JNK inhibitor SP600125 (10 µM) on 4 µM of C-2-induced LC3-II and p-JNK expression changes of BIU87 and EJ cells

Fig 4: Mir30c Is Downregulated and Autophagy Is Induced in Diabetic Mouse Hearts(A) Relative circulating miRNA levels in patients. (B) The correlation between circulating Mir30c levels and fasting glucose. Relative Mir30c expression was determined by qRT-PCR in the heart (C), CMs and NCMs (D), and various organs (E) in 24-week-old animals. (F) Western blotting detection (left) and quantification (middle and right) of the cardiac autophagy-related proteins BECN1 and LC3. ACTB was used as an internal control. (G) Representative autophagic vacuoles from cardiac tissues (arrows) analyzed by transmission electron microscopy (magnification ×38,000). Data are expressed as mean ± SEM (n = 8). For all panels, *p < 0.05 versus C57BL/Ks; **p < 0.01 versus C57BL/Ks. Data are representative of three independent experiments.

Fig 5: NUCKS silencing induces autophagy and apoptosis in gastric cancer cells. a Cleaved Caspase 3 and Cleaved PARP were detected by Western blot analysis after the downregulation of NUCKS. The drug-resistant cells were selected and continued to culture for 3, 6 and 9 days for Western blot analysis. b Detecting the change of gastric cancer cells morphology by light microscopy. c Cells transfected with the GFP-LC3B plasmid after NUCKS knockdown and restoration were examined using fluorescence microscopy (Scale bars, 10 µm). The quantification of LC3B-positive puncta is presented as a histogram (*P < 0.05, **P < 0.01, ***P < 0.001). d Immunofluorescence staining with a LC3B antibody was performed to confirm the induction of autophagy after NUCKS downregulation and upregulation. Representative LC3B-positive cells are shown. e The level of autophagy was evaluated by p62 and LC3B expression, as determined using Western blot analysis after NUCKS knockdown and restoration. f CQ blocked shNUCKS-induced autophagy. Cells were pretreated with 50 µM CQ and analyzed at indicated time points by Western blot