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. 2018 Nov 19;9(12):1149. doi: 10.1038/s41419-018-1187-4

Fig. 4. H19 activates cancer autophagy to mediate 5-Fu resistance.

Fig. 4

a The basic levels of LC3 protein in CRC cell lines were detected by western blotting assays. b Western blotting was used to detect LC-3I, LC-3II, and p62 expression in HCT8 and HCT8Fu cells transfected with pcDNA3.1 or pcDNA-H19 and si-NC or si-H19, respectively. c, d Autophagosomes were observed by transmission electron microscopy in HCT8Fu and HCT8 cells transfected with si-NC or si-H19 and pcDNA3.1 or pcDNA-H19, respectively. For starvation, indicated cells were cultured in serum free Earle’s balanced salt solution (EBSS) medium as the positive control for observation of Autophagosomes. Bar scale, 2 μm. e, f LC3 aggregation in HCT8Fu cells (e) transfected with si-NC and si-H19 detected by the confocal microscope (Bar scale, 50 μm) and in HCT8 (f) transfected with pcDNA3.1 or pcDNA-H19 observed under the fluorescence microscope. Bar scale, 15 μm. g The cell sensitivity of HCT8 and HCT116 transfected with pcDNA3.1 or pcDNA-H19 to 5-Fu was evaluated using the MTT assay upon exposure to the step-up concentration of 5-Fu with or without CQ administration for 72h. **P<0.01, ***P<0.001. h, j Apoptosis was detected by flow cytometry in HCT8, HCT116 cells transfected with pcDNA3.1 or pcDNA-H19. The cells were treated with PBS or CQ. i, k Columns are the average of three independent experiments. Data are presented as mean±SD from three independent experiments. **P<0.01, ***P<0.001