Fig. 6

Signal transduction coupled to G protein is altered in mutant ADGRL2 amniocytes. Intracellular calcium levels were monitored by microfluorimetry using the ratiometric Fura-2 AM calcium probe and results expressed as mean fluorescence intensity (MFI). a α-latrotoxin (1 nM) was applied to wild-type (Wt) and mutant (Mt) cultured amniocytes under extracellular chelated-calcium conditions (EDTA 4 mM). Three minutes after α-latrotoxin administration, cells were perfused without EDTA to restore extracellular calcium levels. b α-latrotoxin (1 nM) was applied to Wt amniocytes under chelated-calcium conditions (EDTA 4 mM). Cultured cells were pre-incubated or not with the phospholipase C inhibitor U73122 (10 μM). Three minutes after α-latrotoxin administration, cells were perfused without EDTA to restore extracellular calcium levels. c α-latrotoxin (1 nM) was applied to Mt amniocytes under chelated-calcium conditions (EDTA 4 mM). Cultured cells were pre-incubated or not with the phospholipase C inhibitor U73122 (10 μM). Three minutes after α-latrotoxin administration, cells were perfused without EDTA to restore extracellular calcium levels. d Quantification and statistical analysis of intracellular calcium levels from the early and late phases in response to α-latrotoxin stimulation. Areas under the curves (AUC) were expressed in arbitrary units (AU). Each value represents the mean (±S.E.M.) of 30 cells. *, p < 0.05; **, p < 0.01, vs Wt amniocytes using one-way ANOVA test