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. 2013 May 31;8(5):e65163. doi: 10.1371/journal.pone.0065163

Figure 5. Smad3-dependent transcriptional activation is regulated by phosphorylation of 14-3-3σ at Ser69 and Ser74.

Figure 5

(A) CAGA (12)-luc reporter activation upon expression of 14-3-3σ constructs and Smad3, * - p value<0.05 (B) DNA precipitation assay shows the recruitment of Smad3 to the CAGA element in the present of wild-type and mutant 14-3-3σ proteins. Expression controls for Smad3 and 14-3-3σ constructs are shown in two lower panels. Densitometry analysis of the protein precipitation is shown in the accompanying graphs. (C, D) TGFβ dependent and Smad3 governed transcriptional activation of PAI-1 and COL7A1 genes. For gene expression analysis, MCF10A cells were stably transfected with the constructs expressing wild type or mutated 14-3-3σ protein and treated with TGFβ1 at concentration of 5 ng/ml for 12 hours. The data shown correspond to a representative experiment out of three performed. (E) Schematic presentation of the restrictive role of 14-3-3σ phosphorylation on TGFβ1/Smad3-dependent transcription.