Fig. 5

ASB9 overexpression inhibits GCs proliferation. A The overexpression efficiency of ASB9 was detected using RT-qPCR. Data are expressed as mean ± SEM (n = 6), ^****P < 0.0001. B Western blotting reveals the expression levels of ASB9. C Quantitative statistics of ASB9. Data are expressed as mean ± SEM (n = 3), ^**P < 0.01. D Flow cytometry determines cell percentages in different cell cycle phases. E Cell cycle analysis statistical results. Data are expressed as mean ± SEM (n = 4), ^*P < 0.05. F EdU staining was used to detect the number of proliferating cells. RED, EdU-positive cells; BLUE, Hoechst staining for total nuclei. Data are expressed as mean ± SEM (n = 3), ^*P < 0.05. G CCK-8 assay detecting cell viability at 24 h after transfection. Data are expressed as mean ± SEM (n = 16), ^**P < 0.01. H RT-qPCR analysis of proliferation-related genes, including CCNB1, CCND1, CCNE1, CDK1, and CDK4. Data are expressed as mean ± SEM (n = 6), ^*P < 0.05, ^**P < 0.01. I Western blot analysis of proliferation-related gene protein level (ASB9, CCNB1, CCNE1, CDK4, and CDKN1A). J Quantifying the western blot analysis of ASB9, CCNB1, CCNE1, CDK4, and CDKN1A. Data are expressed as mean ± SEM (n = 3), ^*P < 0.05