Figure 2.

NFATc3-dependent Twist2 expression is repressed by JNK activity. (a) Thymocytes of DO-TCR-Tg mice were isolated at 3 h after i.p. injection with either PBS or OVAp (100 μg). Twist2 expression are presented relative to PBS-injected DO-TCR-Tg mice (error bars, ± S.E.M., n=4). (b) Thymocytes from Tcrα^−/− mice were treated with P0.1/I200 or P10/I500 for the indicated time period (0, 1, 3, 6, and 9 h). Twist2 expression was measured by qPCR (error bars, ±S.E.M.). (c) 16610D9 cells were treated with I0/P0 (non), I200, P10/I200, or P10/I200+SP600125 for 3 h. Cells were stained with DAPI (blue) and anti-NFATc3 antibody, which was followed by staining with anti-rabbit-TRITC (red) antibody. (d) ChIP assay was performed by using anti-NFATc3 antibody after stimulation of 16610D9 cells with P0.1/I200 and P10/I500 for 0, 1, 2, and 3 h. The eluted supernatants as well as the Input extracts were PCR-amplified with primers for Twist2 promoter in the −185~+63 region