Skip to main content
. Author manuscript; available in PMC: 2022 May 5.
Published in final edited form as: J Vasc Res. 2021 May 5;58(5):277–285. doi: 10.1159/000516044

Figure 2. RapGEF1 silencing eliminates elevated Pi-induced ERK1/2 phosphorylation.

Figure 2.

Primary murine VSMCs were transfected with either RapGEF1 siRNA (siRapGEF1) or negative control siRNA (siNegative). A) RapGEF1 RNA was quantified with qRT-PCR on day 2, 3, and 4 after siRNA transfection. Immunocytochemistry probing for RapGEF1 (green) and was used to visualize RapGEF1 protein in B) siNegative and C) siRapGEF1 VSMCs (scale bar = 100μm). D) 10 cells/image were quantified for total cell fluorescence to show RapGEF1 protein depletion in siRNA treated VSMCs. E) Both siNegative and siRapGEF1 VSMCs were incubated in media containing either 0.5mM, 1.0mM, or 3.0mM Pi for 15 minutes ERK1/2 phosphorylation was visualized by western blot (representative image shown). F) Quantification of six independent samples was performed by densitometry analysis. (two-way ANOVA post-hoc Sidak in A, student’s t-test in D, one-way ANOVA post-hoc Tukey in F: ns = p > 0.05, * = p < 0.05, ** = p < 0.01, *** = p < 0.001, **** = p < 0.0001)