Fig. 3

PHF5A regulates the protein stability of FOS through SKP2-mediated ubiquitination. A Prime View Human Gene Expression Array was performed to identify DEGs between shCtrl and shPHF5A groups of MGC-803 cells. In the heat map of cluster analysis, each column represents a sample and each row represents a differential gene. The red indicates that the gene expression is relatively up-regulated, the green indicates that the gene expression is relatively down-regulated, the black indicates that the gene expression is not significantly changed, and the gray indicates that the signal strength of the gene is not detected. B, C The most significant DEGs in MGC-803 cells with PHF5A knockdown were screened by qRT-PCR and western blot analysis. D The effect of FOS, PTEN, PRKCZ knockdown on proliferation was detected using Celigo cell counting assay. E The E3 ubiquitin ligase affecting FOS protein stability was predicted by bioinformatics analysis. F, G The protein stability of FOS in MGC-803 cells after PHF5A knockdown and SKP2 overexpression was examined. H, I After treatment with MG-132, the protein stability of FOS in MGC-803 cells after PHF5A knockdown and SKP2 overexpression was examined. J, K The lysates of MGC-803 cells after PHF5A knockdown and SKP2 overexpression were immunoprecipitated and WB was performed to examine the ubiquitination of FOS. L Co-IP analysis of interaction of PHF5A and SKP2 in MGC-803 cells. The presented results were representative of experiments repeated at least three times. Data was represented as mean ± SD. ***P < 0.001