Microarray analysis of EZH2 knockout HaCat cell lines

Microarray analysis was performed to screen differentially expressed genes between EZH2 knockout HaCat cell lines and Control HaCat cell lines. From the data of microarray analysis, 344 differentially expressed genes were found between EZH2-shRNA and control group using 1.3-fold cutoff. Of these genes, 254 genes were up-regulated in EZH2-shRNA group and 90 genes were down-regulated.

Methods
Lentiviral vectors were packaged with shRNA targeting EZH2 or full-length Ezh2 gene by Sangon biotech (Shanghai, China). HaCat cells were infected with these lentiviral vectors according to the manufacturer’s instructions.Then stable EZH2 knockdown HaCat cell lines were constructed using puromycin.Three stable EZH2 knockdown HaCat cell lines and three control HaCat cell lines were used for microarray analysis. Total RNA was extracted using TRIzol reagent (Thermo Scientific). Microarrays were performed using Affymetrix GeneChip PrimeView™ Human Gene Expression Array chips (Affymetrix, Santa Clara, CA). The resulting data were analyzed by CapitalBio Technology Company (Beijing, China). A 1.3-fold change cut-off (increased to more than 1.3 times or decreased to less than 0.76 times of the control group) was set to screen differentially expressed genes.