Fig 2: Day 60 DKO cerebral organoids have deficient neuronal marker expression. a Day 60 DKO cerebral organoids derived from hiPSCs demonstrate reduced expression of PAX6, β-tubulin, and CTIP2. Scale bar = 100 µm. b Day 60 DKO cerebral organoids have reduced expression of SOX2, SATB2, and TBR1. Scale bar = 100 µm. c Day 60 DKO cerebral organoids have reduced expression to SOX2, SATB2, S100. Scale bar = 100 µm. All day 60 experiments were conducted after three independent differentiations.

Fig 3: Differentiation of BAX/BAK DKO into hNPCs and validation of cell death phenotype. a Diagram of hiPSCs to hNPCs differentiation protocol based on inhibiting BMP and TGFB pathways. b Derived hNPCs were stained for neural progenitor markers PAX6, SOX2, and Nestin to validate. hiPSCs were also stained for PAX6 and SOX2 to validate. c hNPCs DKO clones have decreased cleaved caspase-3 activity due to loss of BAX and BAK with 24-h etoposide exposure. d hNPCs DKO clones demonstrate a decrease in CC3 compared to controls with 24-h CCCP exposure. Three independent experiments were conducted for etoposide and CCCP exposures. Error bars: SEM.

Fig 4: Day 30 DKO cerebral organoids show cortical disorganization and loss of cortical plate markers. a Differentiation protocol from hiPSCs to day 30 cerebral organoids. b Day 30 DKO cerebral organoids derived from hiPSCs demonstrate reduced expression of SOX2, TBR1, and CTIP2. Scale bar = 100 µm. c Day 30 DKO cerebral organoids have reduced expression of PAX6, Reelin, and TBR2. Scale bar = 100 µm. All day 30 experiments were conducted after three independent differentiations.