Fig. 2

Loss of MECP2 impairs neurogenesis during brain organoid development. A Schematic procedure of culturing brain organoids derived from hESCs. B Representative images of immunofluorescence staining for KI67, TUNEL, pH3S10, TUJ1 and PAX6 in organoids at day 60 of culture. C–F Percentages of KI67 (C), pH3S10 (D), PAX6 (E), TUNEL (F) positive cells among DAPI⁺ cells in organoids at day 60 of culture. n = 3 independent experiments. G Ratio of TUJ1⁺ layer to the whole thickness of cortex-like structure in organoids after 60 days of organoids culture. H Representative traces of mEPSC recordings in neurons of MECP2-KO and PX330 organoids at day 120 of culture. Scale bars, 5 pA and 1 s. I Quantification of mEPSC frequency in PX330 and MECP2-KO organoids. n = 11–19 neurons from 4 organoids per group. J Quantification of mean mEPSC amplitude in PX330 and MECP2-KO organoids. n = 11–19 neurons from 4 organoids per group. *p < 0.05, **p < 0.01, ***p < 0.001. Data are presented as mean ± SEM; Two-tailed Student’s t-test