Fig. 2.

(A) [Pyr¹]apelin-13 stimulated ERK activation in mAPJ-HEK293 at the time points indicated and (B) with the indicated concentrations of [Pyr¹]apelin-13 for 5 min. Cells were fixed, stained, and imaged for determination of whole-cell ppERK1/2 using anti ppERK1/2 antibody, and total ERK1/2 measured with anti-ERK1/2 antibody. The value determined with no primary antibody present was designated as background and was subtracted from raw data to give arbitrary fluorescence units (AFU) and then normalized to a percentage of vehicle control. Data shown are mean ± SEM, of at least three separate experiments, each with triplicate wells and triplicate fields within wells. *p < 0.05 and **p < 0.01 comparing stimulations to basal conditions, analysed by one-way ANOVA and Dunnett's multiple comparison post hoc tests.