Fig. 6: K668 methylation is required for NFAT5 induced limited TMZ efficacy by enhanced MGMT transcription.
From: Lysine methylation promotes NFAT5 activation and determines temozolomide efficacy in glioblastoma
a CCK-8 assay analysis revealed the effect of NFAT5 K668 methylation on TMZ efficacy at the indicated concentrations for 72 h. b Colony formation assay in cells expressing NFAT5 WT or K668R with TMZ treatment. c The effect of NFAT5 on MGMT mRNA levels in U87/EGFR and U251 cells. d The results of the dual-luciferase reporter assay revealed that MGMT was a direct transcriptional target of NFAT5. e ChIP assay demonstrated that NFAT5 K668R mutation reduced NFAT5 binding affinity to MGMT promoter, compare to the NFAT5 WT cells. f CHIP assay revealed that EZH2 knockdown by shRNA decreased the binding affinity of NFAT5 to the promoter of MGMT. g The mRNA and protein levels (h) of MGMT in U87/EGFR and U251 cells expressing NFAT5 WT or K668R mutation. a–h n = 3 independent experiments. Significance was calculated by (b, c, g) unpaired Student’s t test; (d, e, f) by one-way ANOVA with LSD-t. Data were presented as mean ± standard deviation. Marker unit for Western blots is kDa. Marker size for CHIP is bp. Source data are provided as a Source Data file.
