PRMT5 imposed a dual repression on DDIT3 transcription to promote the malignancy of hepatocellular carcinoma

Figure 2 Inhibition of PRMT5 induced cell apoptosis and increased DDIT3 expression in hepatocellular carcinoma cells. A-D: Cell apoptosis was examined in knockdown and PRMT5-inhibited hepatocellular carcinoma (HCC) cells by flow cytometry analysis; E: Heatmaps showed that knockdown PRMT5 upregulated the messenger RNA (mRNA) level of DDIT3 from RNA-sequencing cohort (GSE168745); F and G: The expression of DDIT3 mRNA was evaluated in PRMT5 overexpression and knockdown HCC cells; H and I: DDIT3 mRNA expression was assessed in HCC cells treated with HLCL-61 at different times or different concentrations of HLCL-61; J: Western blot was performed to examined the expression of DDIT3 and PRMT5 protein in stable cell lines; K: DDIT3 protein level was detected in HCC cells treated with HLCL-61 (8 μM) at 0, 12, 24, 36, 48 hours; L: The PRMT5 and DDIT3 expression were examined in paired HCC tissues and their adjacent normal tissues; M: Based on the results of western blot, the correlation between PRMT5 and DDIT3 expression was analyzed by statistical analysis. Data are presented as the mean ± SD. ^aP < 0.05. ^bP < 0.01. ^cP < 0.001. NS: Not significant; Scr: Scramble; FITC: Fluorescein isothiocyanate; PI: Propidium iodide; mRNA: Messenger RNA; N: Normal; T: Tumor; shPRMT5: Short hairpin RNA of PRMT5.