Fig. 4
SHANK3 mRNA puncta analysis in control hiPSC neurons over developmental time line. a, b SHANK3 mRNA identified through smFISH. Top to bottom; bright-field (BF) image of representative hiPSC-derived neuron, SHANK3 mRNA smFISH image, composite smFISH and BF image, and SHANK3 mRNA puncta selected through quantification analysis. Images for (a′) 1 day in vitro (DIV) (left; n = 17) and (b′) 99 DIV (right; n = 11). Bar graph represents SHANK3 mRNA puncta per 100 μm2 for (a”) cell body (left) and (b″) processes (right). Error bars indicate standard error of mean. *Unpaired t-test p < 0.0001. c Z-stack demonstrates SHANK3 mRNA located throughout cell body of hiSPC-derived neurons as seen in maximum image projection (MIP). Z-stack images taken 1.0 μm apart. mRNA messenger RNA, SHANK3 SH3 and multiple ankyrin repeat domains 3