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. 2017 May 15;114(22):5683–5688. doi: 10.1073/pnas.1700861114

Fig. S6.

Fig. S6.

Reciprocal regulation of p53 conformation by XAF1 and MT2A. (A) Inverse response of XAF1 and MT2A to increasing doses of CdCl2 and its association with p53 and XIAP expression in U2OS-XAF1 subline cells. IB and semiquantitative RT-PCR assays were carried out to determine p53 traget gene expression after 24 h treatment. (B) A semiquantitative RT-PCR analysis showing MT2A repression of p53 target gene expression induced by cytotoxic metal stress. (C) No detectable effect of XAF1 on p53 conformation and XIAP level in MT2A-depleted cells. PAb1620 and PAb240 antibodies were used to detect WT and mutant-type (under nondenaturating gel conditions) p53 conformation, respectively. Pan-p53 antibody was used to determine total p53 protein level. (D) Opposite effects of XAF1 and MT2A on mRNA expression of p53 target genes. HCT116 cells were transfected with Flag-XAF1 and/or MT2A expression plasmids, and semiquantitative RT-PCR analysis was performed at 48 h after transfection. (E) A comparative analysis of XAF1’s apoptosis-promoting effect in p53+/+ and p53−/− cells exposed to metal stress (CdCl2, 48 h).