Fig. 3

Rsrc1-161aa regulates cell proliferation and cell cycle progression in GC-1 spg cells and interacts with C1qbp. a Upper figure: schematic diagram of linearized Rsrc1-161aa. Green triangle, start codon; orange and blue squares, Rsrc1-161aa sequence; yellow square, HA epitope; red square, stop codon. Lower figure: fluorescence images of Rsrc1^+/+/circRsrc1^−/− GC-1 spg cells stably transfected with plasmids expressing Rsrc1-161aa. Bar, 20 μm. b The cell growth potential was determined based on cell number in wild-type, Rsrc1^+/+/circRsrc1^−/.⁻ and rescue cells. Bar, 400 μm (n = 27). c The effect of Rsrc1-161aa on cell proliferation by CCK-8 assay (n = 5). d The regulatory effect of Rsrc1-161aa on cell cycle progression (n = 3). e Rsrc1-161aa related proteins were identified by mass spectrometry after immunoprecipitation using anti-GFP antibodies and silver staining. f Western blotting of co-immunoprecipitated proteins. g Representative confocal microscopy images of GC-1 spg cells after staining with C1qbp (green) and Mitotracker (red). Bar, 10 μm. h Representative immunofluorescence images of GC-1 spg cells transiently transfected with Rsrc1-161aa-HA with PLA signal for detection of HA–C1qbp interaction (red). Nuclei were stained with DAPI (blue). Bar, 10 μm. All data are means ± SEM, tested using one-way ANOVA with Tukey’s post hoc analysis *P < 0.05; **P < 0.01; ***P < 0.001