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. 2012 Dec 7;288(3):1883–1895. doi: 10.1074/jbc.M112.417881

FIGURE 1.

FIGURE 1.

Induction of JNK1 activity by Netrin-1. A and B, Netrin-1 increased JNK1 activity in the presence of DCC (A) and DSCAM (B). Human full-length JNK1 was co-transfected with either DCC-Myc (A) or DSCAM-Flag (B) in HEK293 cells. JNK1 was immunoprecipitated with anti-JNK and JNK activity performed following the JNK activity assay kit instructions. C and D, time-dependent induction of JNK1 activity by Netrin-1 in HEK293 cells expressing DCC (C) or DSCAM (D). Netrin-1 increased JNK1 activity within 5 min, and this induction was persistent over 20 min. E and F, additive effect of DCC and DSCAM on Netrin-1-induced JNK1 activation. Human full-length JNK1 was co-transfected with either DCC only, DSCAM only, or DCC plus DSCAM in HEK293 cells. Cells were stimulated with purified Netrin-1 (250 ng/ml) for 20 min. F, quantification of the relative induction of JNK1 activity of E. Netrin-1 increased JNK1 activity in HEK293 cells expressing DCC or DSCAM only, and expression of both DCC and DSCAM together led to an even more pronounced Netrin-1-induced JNK1 activity. The relative induction of JNK activity was examined by the ratio of band intensity of Netrin-1 group versus the control. Data are mean ± S.D. from three separate experiments. One-way ANOVA and Fisher's LSD post hoc comparisons were performed. ***, p < 0.001.