Fig. 6

Knockdown of UBE2C affected the GC proliferation and cell cycle progression. A qRT-PCR showed the mRNA expression of UBE2C after KGN were infected with shUBE2C and shNC lentivirus. Among the three shRNA sequences, shUBE2C-1 and shUBE2C-2 showed the significant knockdown efficacy. Data were presented as the means ± SEM from three repeated experiments. B Western blotting analysis showed the protein expression of UBE2C in shUBE2C and shNC cells. Relative expression was normalized to the β-actin and were presented as the means ± SEM from three repeated experiments. C CCK-8 showed that the cell viability was inhibited after depletion of UBE2C in KGN. Data were presented as the means ± SEM from three repeated experiments. D EdU staining assay showed the proliferation of KGN after knockdown of UBE2C. The proliferating cells were stained with EdU (red) and all cells were stained with Hoechst (blue). Scale bar = 100 um. Values were presented as the means ± SEM from three repeated experiments. (E and F) Flow cytometry result showed the cell cycle distribution after UBE2C knockdown in KGN. Data were presented as the means ± SEM from three repeated experiments (*p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001, two-tailed Student’s t-test). UBE2C: ubiquitin conjugating; GCs: granulosa cells; NC: control; CCK-8: Cell Counting Kit-8; EdU: 5‐Ethynyl‐2’‐deoxyuridine; SEM: standard error of mean