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. 2013 Dec 31;8(12):e84063. doi: 10.1371/journal.pone.0084063

Figure 2. Phosphorylation of JNK, the Smad3 linker region (T179 and S208) and the Smad3 C-terminal domain following unilateral ureteral obstruction (UUO) and in sham operated controls.

Figure 2

(A) Immunoprecipitation (IP) of Smad3 followed by Western Blotting (WB) identified phosphorylation of the Smad3 C-terminal domain (p-C-Smad3), phosphorylation of the Smad3 linker region (p-T179) in the UUO kidney. IP of Smad3 also pulled down phosphorylated JNK (p-JNK). Detection of total Smad3 confirms equal efficiency of Smad3 precipitation. The results are quantified in (B). (C) Direct WB of kidney lysates was used to analyse protein levels of TGF-β1, phosphorylation of Smad3 linker region at S208 (p-S208) and total p-JNK. The results are quantified in (D). Data are mean ±SD from groups of 5 mice and analysis by one-way ANOVA with post hoc analysis with Tukey's multiple comparison test. *p<0.05, **p<0.01, ***p<0.001 versus sham operated group.