Figure 3.

CRAC inhibition rescues ligand-induced aberrant calcium signaling in variant cells. (a) TIME-GNAQ^WT or -GNAQ^R183Q were loaded with intracellular calcium probe Fluo-8 and treated for 20 minutes with vehicle or 1 μM CM4620. After treatment, cells were stimulated with thrombin 1U/ml and fluorescence recorded for 300 seconds. The graphs represent an average of three independent experiments. Statistical test comparing GNAQ^R183Q treated by vehicle or CM4620 is described in (b). (b) Means ± SD of AUC calculated from three experiments summarised in Figure 3a. Statistical comparisons were performed by two-tailed paired t-test (n.s. = statistically nonsignificant, ∗P = .0284). (c) TIME recombinant cell lines were assayed for concentration of IP1 after treatment with vehicle or CM4620 0.3μM, 1μM, 3μM, or 10μM. The graph represents the mean ± SD of three independent experiments including 5–6 technical replicates each. Statistical comparison among conditions was performed by two-tailed unpaired t-test (n.s. = nonsignificant, ∗∗P = .0078). AUC, area under curve; IP1, inositol-1-phosphate; ns, nonsignificant; RFU, relative fluorescence unit; TIME, telomerase-immortalised microvascular endothelial; Veh, vehicle; WT, wild type.