Fig. 2

CRNDE induces proteasome ubiquitination-dependent SRSF6 degradation and contributes to autophagy-induced chemoresistance in GC cells. a RNA pull-down assay was used to identify the proteins associated with CRNDE. Biotinylated CRNDE and antisense RNA were incubated with cell extracts, and the associated proteins were resolved by SDS-PAGE. The CRNDE-sense-special bands (arrows) were excised and analyzed by mass spectrometry. b SRSF6 was identified as CRNDE binding protein by mass spectrometry. c RIP assay showed that CRNDE and SRSF6 proteins interact with each other in MGC803 cells. d Western blot confirms the presence of SRSF6 in CRNDE pull-down products. e The effect of CRNDE on the stability of SRSF6. MG-132 eliminates the effect of CRNDE on the stability of SRSF6. (F) CRNDE affects the ubiquitination of SRSF6 in MGC803 cells. g Western blot analyzed the expression of LC3II in MGC803 cells transfected with shSRSF6 plasmid in the presence of CQ. h The chemotherapy resistance of MGC803 cells to oxaliplatin and 5-FU caused by interfering with the expression of CRNDE was counteracted after administration of shSFSF6 in vivo. The tumor volume of each group is shown (n=5). Representative figures are shown. The results are from three independent experiments. Student’s t-test and one-way ANOVA; mean ± SD; the asterisk (**) indicates P < 0.01