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. 2009 Mar 23;106(14):5675–5680. doi: 10.1073/pnas.0809568106

Fig. 3.

Fig. 3.

MKP7 is nitrosylated by nitric oxide. (A) Western blot analysis to detect phosphorylated JNK and total JNK protein in BAECs pretreated with Na3VO4 (10, 100, and 500 μM) and then treated with 50 ng/mL SDF-1α for 10 min. (B) Results of a biotin switch assay on HEK293T cells transfected with Myc-MKP7 and treated with the nitric oxide donor nitrosoglutathione (GSNO) (500 μM) for 10 min at room temperature. (C) Results of an in vitro phosphatase assay using active JNK3 protein as the substrate on lysates of HEK293T cells transfected with Myc-MKP7. (D) Schematic structure of MKP7 mutant constructs. (E) Biotin switch analysis of HEK293T cells transfected with the indicated Flag-tagged MKP7 constructs and then treated with 50 μM GSNO for 10 min at room temperature. (F) Results of an in vitro phosphatase assay using active JNK3 protein as the substrate on HEK293T cells transfected with Flag-tagged MKP7 and then treated with 200 μM GSNO for 10 min. (G) Quantitative analysis of results from in vitro phosphatase assay of MKP7 based on three independent experiments. *, P < 0.05, compared with control cells. #, P < 0.05, compared with the same cells without GSNO treatment. NS, not significant, compared with control cells or the same cells without GSNO treatment, respectively.