Fig. 2.

REST and RCOR gene expression in functional and mature NGN2 iNs. (A) hPSCs were differentiated into functionally mature iNs derived from H9 ESCs and two healthy control iPSC lines (iPSC1 and iPSC2) using NGN2 overexpression. (B) Immunocytochemistry confirmed expression of MAP2 and VGLUT1 at a protein level in neurons derived from all cell lines (n = 3 independent differentiations). Scale bars = 50 μm. (C) Representative current clamp recordings of membrane potential responses of the H9, iPSC1 and iPSC2 cell lines (n = 16, 12 and 16 cells, respectively, from 3 independent differentiations). The stimulation protocols consisted of 1 s long current injections from −100 pA to 100 pA in 10 pA steps. The coloured traces indicate firing activity at rheobase. (D)REST and RCOR gene relative expression levels were analysed via RT-qPCR. Data was calculated as the mean of three housekeeping genes and presented as mean ± SEM. Data was analysed using an ordinary Two-way ANOVA with a Holm-Sidak test to correct for multiple comparisons. *p < 0.05, ***p < 0.001. n = 3 biological replicates, with each data point representing the average of 3 technical replicates.