Fig. 2
From: Increased Ca2+ signaling in NRXN1α+/− neurons derived from ASD induced pluripotent stem cells
Differentiation and validation of the cortical excitation neurons. a–c ICC staining of day-100 neurons with MAP2 (a), SYN1/TUJ1 (b) for neuronal maturity, and with TBR1/CTIP2 for the identity of lower cortical pyramidal neurons (c). d Proportion of layer VI (TBR1+) and layer V/VI (CTIP2+) neurons in the cultures. e High mRNA expression (normalized to GAPDH [dotted line]) of excitatory post-synaptic markers (DLG4, SHANK1, SHANK2, SHANK3, SLC17A7, GRIN1, GRIN2A, GRIA1, GRIA4, GRIK1, GRIK3), neuronal markers (TUBB3, MAP2), upper cortical layers (BRN2, SATB2), and lower layer markers (TBR1, CTIP2) in day-100 cultures of control (black) and NRXN1α+/− (red) neurons. Results shown from two independent cortical neuronal differentiation (f). Representative waterfall traces of spontaneous Ca2+ transients in day 50 (blue) and 100 (pink) cultures from 200 s of recording. Neurons exhibited a significant increase in proportion (%) of spontaneous active cells (g), the frequency (mHz, h), and the amplitude (ΔF/F, i) of Ca2+ transients from day 50 to day 100. Statistical significance (**p < 0.01, ***p < 0.001, ****p < 0.0001) was evaluated using the Mann-Whitney U test). All representative images all from control line 4CCX1