Fig. 3

SHANK3 protein expression increases with developmental age. a SHANK3 protein identified through immunofluorescence labelling. For each panel, left to right: bright-field image (BF) of representative hiPSC-derived neuron, SHANK3 protein immunofluorescence image, composite immunofluorescence and BF image, and SHANK3 protein puncta selected through automated quantification analysis. Images for (a′) 1 day in vitro (DIV) (n = 3), (a′′) 28 DIV (n = 4) and (a′′′) 115 DIV (n = 5). b SHANK3 protein puncta per 100 μm². Error bars indicate standard error of mean. *Kruskal–Wallis test p < 0.005 at 1 DIV vs 115 DIV. c Z-stack demonstrates SHANK3 protein only present on outside and not within cell body of hiSPC-derived neurons as seen in maximum image projection (MIP). Z-stack images taken at 3.0 μm spacing. Yellow dashed lines surround neuronal cell body. SHANK3 SH3 and multiple ankyrin repeat domains 3