Figure 3 | Scientific Reports

Figure 3

From: Identification of PTPN1 as a novel negative regulator of the JNK MAPK pathway using a synthetic screening for pathway-specific phosphatases

Figure 3

PTPN1 negatively regulates the JNK pathway via JNK dephosphorylation. (A–C) Phosphorylation levels of each kinase were examined by immunoblot analysis. PTPN1 was incubated with phosphorylated form of (A) HA-MLK3 and (B) GST-MKK7. (C) Phosphorylated GST-JNK was incubated with PTPN1 (30 min, 37 °C), and then GST-cJun and ATP were added to perform a JNK kinase assay. (D) Flag-PTPN1 was expressed in 293 T cells. Control cells were transfected with an empty vector. After TNFα-stimulation, samples were prepared at the indicated time-points. JNK phosphorylation levels, with or without PTPN1 expression, were examined by immunoblot analysis using an anti-dual phospho-JNK antibody. (E) The density of immunoblotted bands detected using the anti-dual phospho-JNK antibody is plotted. (*p < 0.05) All experiments were performed at least three times.

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