Effect of thiazide on RasGRP1-mediated inhibition of the PTH effect. A: TRPV5 activity with administration of 100 nM PTH for 15 min in control cells and mDCT15 cells with reduced RasGRP1 expression (RG1). Cells were treated with 100 nM PTH for 15 min with and without thiazide diuretic (T). n = 4. *P < 0.01 compared with control; #P < 0.01 compared with RG; @P < 0.01 compared with RG + P. Solid circles indicate means, shaded circles indicate individual data points, and bars represent SEs. B: representation blot showing TRPV5 surface and total expression for control (C), thiazide-treated (T), PTH-treated (P), thiazide and PTH-treated (T + P), RasGRP1 knockdown (R), RasGRP1 knockdown treated with thiazide (R + T), RasGRP1 knockdown treated with PTH (R + P), and RasGRP1 knockdown treated with thiazide and PTH (R + P + T) cells (n = 4).