Figure 3
From: Elabela-Apelin Receptor Signaling Pathway is Functional in Mammalian Systems
Down-stream signaling of APJ activation by ELA.
(A). Suppression of cAMP production by ELA. CHO cells expressing APJ were treated with forskolin (10 μM) and ELA at the indicated concentrations. The amount of cAMP was measured by TR-FRET. Higher TR-FRET ratio indicates less cAMP production. (B). Phosphorylation of ERK1/2 by ELA. Upper panel: APJ-expressing CHO cells were treated with ELA (1 μΜ) at the indicated times and lysed for Western blot analysis by antibodies of phosphorylated form of ERK (pERK1/2) and unphosphorylated ERK1/2 (ERK1/2). (B). Dose response of ERK phosphorylation by ELA. APJ-expressing CHO cells were treated with ELA at the indicated concentrations and assayed for HTRF ratio by TR-HTRF. (C). Intracellular calcium mobilization by ELA. CHO cells expressing human APJ were treated with ELA at the indicated concentration and calcium mobilization was monitored by fluorescent calcium dye Fluo8 for up to 100 seconds (upper panel). Lower panel: calcium response to ELA in APJ-expressing cells vs. vector control cells. Values are mean ± SEM (n = 3–5).
