Fig. 2. Lithocholic acid (LCA)-induced response is specifically mediated by Mrgpra1. (A) The LCA-induced intracellular calcium increase (n=1,342 cells) was suppressed by pretreatment with 1 µM QWF (n=1,767 cells) in Mrgpra1-expressing HEK293T cells (Mrgpra1-HEK293T). (B) A comparison of relative responses between LCA vs. QWF pretreatment. ***p<0.001. (C) Representative fluorescent images of Mrgpra1-HEK293T at initial (0 s) and peak time points for QWF pretreatment before LCA application. Scale bars=100 µm. (D) LCA failed to evoke an increase in intracellular calcium levels in calcium-free media (n=3,900 cells). (E-G) Responses from various inhibitors on LCA treatment in Mrgpra1-HEK293T. LCA-induced responses were significantly suppressed by SKF96365 (E; a non-specific TRPC blocker, 1028 cells), U73122 (F; an inhibitor of PLC, n=969 cells), and YM254890 (G; a Gαq inhibitor, n=914 cells). (H) A summary of relative responses of pretreatment effects with various inhibitors in the Mrgpra1-HEK293T. ***p<0.001.
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