mNgn2-induced neurons and GFP-NPCs at two and three weeks of culture were immunostained with SYN1 and MAP2AB as described in Sections 2.3.1 and 2.3.2. The images were acquired (five images each from three biological replicates per condition and cell line) using a confocal microscope (LSM 780, Zeiss) at 400× total magnification. After uniform thresholding of all SYN1 and MAP2AB images, SYN1 puncta were counted, and the MAP2AB-positive area of the thresholded images was measured using NIH Image J. Total SYN1 puncta per image were divided by that image’s respective MAP2AB-positive area in order to calculate SYN1 puncti normalized to MAP2AB levels.
