Fig. 2.

Effect of cGMP, KT5823, and H8 on TRPC3-mediated store-operated Ca²⁺ influx in PKG-HEK cells. (A) Store-operated Ca²⁺ influx in TRPC3-transfected PKG-HEK cells. Shown is the mean ± SE (n = 14 experiments). (B) Store-operated Ca²⁺ influx in PKG-HEK cells that were transfected with empty vector pcDNA6. Shown is the mean ± SE (n = 14 experiments). Cells were depleted with 4 μM thapsigargin in 0 Ca²⁺-PSS for 15 min. 8-BrcGMP (2 mM) with/without KT5823 (1 μM) or H8 (10 μM) was introduced 5 min before the experiments. At the time indicated by the arrow, the media were changed to 0.5 Ca²⁺-PSS. (C) Subtraction of corresponding data in B from A yields the components that are attributed only to the overexpressed TRPC3. (D Upper) Immunoblots with antibody against phosphorylated VASP (16C2) in PKG-HEK cells. (Lower) Immunoblots with antibody against total VASP (M4) in PKG-HEK cells. Cells were treated with 8-BrcGMP (2 mM) in the absence or presence of KT5823 (1 μM) or H8 (10 μM) for 5 min. Control had no treatment. Only one main VASP band of 50 kDa was consistently observed. Another VASP band of 46 kDa (not shown) was very weak and was observed only occasionally.