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. 2016 Oct 6;6:34543. doi: 10.1038/srep34543

Figure 2. c-Jun Ser73 phosphorylation is essential for ERK-dependent demethylation and activation of Zta promoter.

Figure 2

(a) B95-8 cells were pretreated with U0126 (20 μM) for 2 h and then treated with TPA for 1, 3, 6, 12, and 24 h, respectively. Whole-cell lysates were isolated and the levels of phosphorylated ERK (p-ERK), phosphorylated c-Jun (p-c-Jun), phosphorylated c-Fos (p-c-Fos), ERK, c-Jun, c-Fos, Zta, and GAPDH proteins were determined by Western blotting using corresponding antibodies. (b) B95-8 cells were pretreated with U0126 (20 μM) for 2 h and then treated with TPA for 15 and 90 min. Whole-cell lysates were isolated and the levels of phosphorylated c-Jun (p-c-Jun) and GAPDH proteins were determined by Western blotting. (c) B95-8 cells were treated with TPA for 0.5, 1, 3, 6 and 9 h. Total RNAs were isolated from the cells and the level of Zta mRNA was analyzed by qPCR. *P < 0.05, **P < 0.01 and ***P < 0.001.