Figure 3.

H2AFJ upregulation desensitizes GBM cells to TMZ treatment. (A) RT-PCR analysis of H2AFJ and GAPDH expression in different GBM cell lines. (B) Scatter plots for the correlation between H2AFJ expression and TMZ IC₅₀ concentrations in various GBM cells. (C) Cell viability of the detected GBM cells after 24-h exposure to radiation at 8 Gy. The count of remaining viable cells after radiation treatment was normalized with the cell number of the untreated group in each detected GBM cell line. The data from three independent experiments are presented as the mean ± SEM. (D) Scatter plots for the correlation between H2AFJ mRNA levels and cell viability of the detected GBM cells posttreatment with radiation at 8 Gy for 24 h. A non-parametric Spearman correlation test was used to estimate statistical significance in B and D. (E,F) Scatchart plots for the cell viability of D54MG cells (E) with or without H2AFJ knockdown (insert) and T98G cells (F) with or without H2AFJ overexpression (insert) after TMZ treatment for four days at the indicated TMZ concentrations. In A, E and F, GAPDH was used as an internal control for RT-PCR experiments. (G) Kaplan–Meier analysis of overall survival probability associated with H2AFJ gene expression in MGMT-unmethylated and -methylated GBM patients undergoing radiation therapy or standard TMZ/radiation therapy. (H) Scatter plots for H2AFJ transcripts and time to a new tumor event after radiation or TMZ therapy in GBM patients who were diagnosed to be wild-type IDH and non-G-CIMP. The statistical significance was estimated by Pearson’s correlation test. The symbol “n.s.” denotes not significant.