Fig. 5

Inhibition of neddylation enhances cell cycle signaling pathway. DMSO and MLN4924 were introduced into the granulosa cell culture medium at a concentration of 1 µM. After a 24 h incubation period, the cells were collected for further analysis. Subsequently, RNA and protein of the cells were extracted respectively. A Changes in the expression of apoptosis marker genes Bax, Caspase3, p53 and Bcl2 in GCs after MLN4924 treatment, N = 3 for each group. B Changes in the expression of proliferation marker genes (PCNA, CDK4, CDK6, and CCND1) in GCs after MLN4924 treatment. C The protein levels of cell apoptosis and cell proliferation maker genes were assessed using Western blot analysis. D Quantitative results of protein expression. The intensities of protein bands were quantified using Image J. β-actin was used as the internal control. All the experiments were repeated at least 3 times. Student’s t test were used to compare the differences between groups. Data are shown as mean ± SDs, and the statistical analysis results were visualized using GraphPad Prism5. *** represents P < 0.001, ** represents P < 0.01, * represents P < 0.05, and ns represents non-significant