Fig. 3: The NDD-associated AIRIMV190G variant causes growth defects at a specific stage of cerebral organoid development.
From: A programmed decline in ribosome levels governs human early neurodevelopment
a, Schematic explaining the origin of the variant samples used in this study. b, Schematic of the timeline for generating brain organoids from pluripotent stem cells. c, Bright-field images of control and mutant (V190G) organoids at days 5, 10 and 15. Black arrows indicate NE buds, which seem less organized and elongated in the mutant at day 15. Scale bar, 1 mm. d, Quantification of bright-field images at days 5 (left), 10 (middle) and 15 (right) show that control neural tissue is enlarged relative to mutant (V190G); ****P < 0.0001, unpaired two-sided t-test with Welch’s correction, day 5, n = 73 control EBs 62 mutant EBs from three independent batches; day 10, n = 77 control organoids 84 mutant organoids from four independent batches; day 15, n = 46 control organoids 51 mutant organoids from three independent batches. Error bars show s.e.m. P = 0.8776 (day 5), P = 0.1128 (day 10), P = 2.515 × 10−11 (day 15). NS, not significant. e, Representative images showing the proliferation marker pH3 (grey) and Ki67 (green) signal in control (WT) and mutant (V190G) day 15 organoids. Scale bar, 50 μm. f, Representative images showing TUNEL labelling in control and mutant (V190G) day 15 organoids. Mutant organoids exhibit a higher TUNEL signal. Scale bar, 50 μm. g, Quantification of pH3 signal of individual NE buds at day 15 control (WT) and mutant (V190G) organoids. Unpaired two-sided t-test with Welch’s correction. n = 12 control and 11 mutant imaged regions from six organoids from two independent batches. Error bars show s.e.m. h, Quantification of TUNEL signal of individual NE bud showing that mutant organoids exhibit a higher TUNEL signal. TUNEL counts were normalized to the area of the imaged bud. **P < 0.01, unpaired two-sided t-test with Welch’s correction, P = 0.0022, n = 6 control n = 6 mutant imaged regions from two independent batches. Error bars show s.e.m. The image in a was made using BioRender.
