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. 2016 Sep 12;113(39):E5731–E5740. doi: 10.1073/pnas.1611532113

Fig. S5.

Fig. S5.

AKT phosphorylation is not affected by ectopic expression of Spry2 in mammary fibroblasts. (A–D) Western blot analysis of phosphorylation status of AKT in control and Spry2-GOF fibroblasts in response to AREG (A and B) or TGFα stimulation (C and D). Fibroblasts were serum-starved, treated with 5 nM AREG or TGFα for the indicated durations, and lysed, and on a single blot, P-AKT (Ser473), AKT, and β-actin signals were detected. (B and D) Quantitative comparison of AKT phosphorylation, normalized to total AKT. Data are mean ± SD (n = 2–3). Statistical analysis was performed using two-way ANOVA and revealed no significant difference in P-AKT between control and Spry2-GOF fibroblasts. (E) Relative expression of Spry2 in control and Fsp-Cre;Spry2-GOF fibroblasts, normalized to Actb. The plot shows mean ± SD. Statistical analysis was performed using paired t test; *P < 0.05. (F) Relative expression of Egr1 and Fos in control and Fsp-Cre;Spry2-GOF fibroblasts in response to AREG or TGFα stimulation, normalized to Actb and compared with their expression in mock-treated cells (indicated by dotted line). The plots show mean ± SD. Statistical analysis was performed using two-way ANOVA; *P < 0.05; ***P < 0.001.