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. 2015 Oct 7;5:14838. doi: 10.1038/srep14838

Figure 1. XAF1 expression is induced by either TNF-α or IFN-α in demethylating conditions.

Figure 1

(a) MCF-7 cells were pre-treated with 5-Aza-2′-deoxycytidine (5 μM) and Trichostatin-A (0.2 μM) for 3 days before stimulation with TNF-α (20 ng/mL) for 24 h. Quantitative PCR (qPCR) analysis of XAF1 and CTCF mRNA expression was performed. HPRT mRNA was used as loading control. Results are presented in terms of fold change. The means from three independent experiments were plotted with ±SEM, *P < 0.05. (b) MCF-7 cells were treated as shown in (a). Using a specific antibody, XAF1 was immunoprecipitated from equal quantities of total extracted proteins for each condition. XAF1, CTCF and GAPDH protein levels were measured by Western blot. (c) MCF-7 cells were pre-treated as in (a) before stimulation with IFN-α at the indicated concentrations. mRNA expression of both XAF1 and CTCF was analysed by qPCR after normalizing with HPRT mRNA. The mean from three independent replicates were plotted with +SEM, *P < 0.05. (d) MCF-7 cells were pre-treated and stimulated as shown in (c). Western blot analysis was performed as shown in (b). 5-Aza-2′-deoxycytidine (5-A-DC); Trichostatin-A (TSA).