Sprouty4 negatively regulates ERK/MAPK signaling and the transition from in situ to invasive breast ductal carcinoma
Fig 3
Overexpression of Sprouty4 in MCF10.CA1d cells suppresses ERK/MAPK phosphorylation.
(A) MCF10.CA1d control and Sprouty4 overexpressing cells (MCF10.CA1d/S1 and MCF10.CA1d/S2) grown in 2D culture were serum starved for 24 hours, then treated with DMSO (vehicle) or 200 nM phorbol 12-myristate 13-acetate for 10 minutes. Membranes were immunoblotted for Sprouty4, phosphorylated ERK1/2 (pERK1/2) as well as total ERK1/2, and β-tubulin. Expression levels are representative of three independent experiments. (B) ERK1/2 phosphorylation was quantified using densitometry and plotted using Graphpad Prism. One-way ANOVA employing a repeated measures test and Bonferroni correction was used to assess significance (α = 0.01, 99% CI). Whiskers represent the minimum and maximum values and bars represent the median. Differences were observed under both unstimulated (** p < 0.01) and stimulated (*** p < 0.001) conditions.