After sorting all subjects by their Ca2+ signaling normalized to the reference cell line (as was done for Fig. 2), we generated an ROC curve by plotting sensitivity (true positive rate) against 1-specificity (false positive rate) at each test value for individuals with syndromic ASD (FXS, TSC1 and TSC2, Rett and PWS) as shown in Fig. 3a. This evidence concerns the gene TSC1 and Prader-Willi syndrome.