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. 2013 Jan 29;3:1160. doi: 10.1038/srep01160

Figure 2. TMZ-induced TAp63 suppresses MYC expression in human GBM cells.

Figure 2

(a) TAp63 and MYC mRNA levels in GBM cells with increasing TMZ concentrations, 24 h. (b) Immunoblot of TAp63 and MYC from GBM cells treated with increasing concentrations of TMZ, 24 h. (c) RT-PCR analyses of relative MYC expression in U87MG and YH-13 cells following siTAp63 transfection, normalised to ACTB mRNA. (d) Analysis of MYC expression in YH-13 cells transfected with siControl or siTAp63 by western blotting. (e) RT-PCR analyses showing MYC suppression after TAp63α overexpression in GBM cell lines. (f) Positions of PCR primer sets R1, R2, R3 and R4 for the chromatin immunoprecipitation (ChIP) assays. (g) Identification of the TAp63-binding region in the MYC promoter by ChIP assays. YH-13 cells were transfected with or without indicated siRNAs. Genomic DNA was amplified by PCR using the indicated primers. (h) Semi-quantitative PCR and (i) quantitative PCR of ChIP assays showing endogenous TAp63 recruitment onto the MYC promoter after 24 h TMZ treatment, 150 μM. *P < 0.005 (two-tailed t-test). (j) Luciferase activity of MYC reporters after lentiviral TAp63α or GFP infection of U87MS cells. Data shown as the fold change in the luciferase activity compared with control cells.