On day 12 of differentiation, iPSNs were plated in 24 well optical bottom plates (Cellvis). At day 32 of differentiation, iPSNs were fixed and immunostained as previously described [3 (link)]. Antibodies for immunostaining are as follows: 1:500 Rabbit Anti-Lamin B1 (Abcam ab16048), 1:1000 Guinea Pig Anti-Map2 (Synaptic Systems 188,004), 1:1000 Goat Anti-Rabbit Alexa 488 (Invitrogen A11034), 1:1000 Goat Anti-Guinea Pig Alexa 647 (Invitrogen A21450). iPSNs were imaged using a Zeiss LSM 800 confocal microscope. All images were acquired using identical imaging parameters (e.g. laser power, gain). Unless otherwise indicated, images presented are maximum intensity projections generated in Zeiss Zen Blue 2.3.
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