Fig. 4

Inhibition of neddylation negatively regulates the PPAR signal pathway. DMSO and MLN4924 were added to the granulosa cell culture medium at a concentration of 1 µM. After 24 h of incubation, the cells were collected, and both RNA and protein were subsequently extracted separately (N = 3 for each group). A RT-qPCR results illustrate changes in the transcriptional levels of genes downstream of the PPAR signal pathway and genes related to lipid synthesis. B Western blot analysis was used to assess the protein levels of target genes of the PPAR signaling pathway and lipid synthesis-related genes. PPARg consists of two subtypes, PPARγ1 and PPARγ2. The intensities of protein bands were quantified using Image J C. β-actin served as the internal control. All the experiments were replicated at least 3 times. Student’s t test was employed to compare the differences between groups. Data are presented 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