Figure 1.

Generation, characterization, pluripotency validation and neuronal differentiation of hiPSC colonies from Ctrl and A-T patients. Representative images of newly formed hiPSC colonies before picking (a, upper panel) and after picking (a, bottom panel). The hiPSC colonies were characterized by western blot (a, right) to evaluate the expression of ATM protein and the pluripotency marker Oct 3/4 and were compared with primary fibroblasts and MEF feeders. Vinculin was used as a loading control. In b, hiPSC colonies were labeled to visualize the expression of the pluripotency markers Oct 3/4, SSEA4 and TRA-1-81. Nuclei were counterstained with DAPI (blue). In c, Ctrl and A-T hiPSCs were differentiated in vitro into the three germ layers. After 20 days of differentiation, cells were labeled with antibodies specific for α-SMA (mesoderm marker), Sox17 (endoderm marker) and β-tubulin III (ectoderm marker). Nuclei were counterstained with DAPI (blue). The ability of hiPSCs to generate neuronal cells was confirmed through the formation of floating EBs and rosette formation. Representative images for each differentiation step are shown in d