Fig. 4

Differential effects of APOE deficiency on gene expression related to cholesterol biosynthesis in excitatory neurons and radial glia. a Neurons were isolated from the cerebral organoids through FACS using the cell surface marker CD90. Representative fluorescence microscopy images of the isolated neurons stained with antibodies against Tuj1. Scale bar: 20 μm. b The mRNA levels of selective cholesterol biosynthesis genes in the isolated neurons were quantified by RT-qPCR (n = 4 wells/genotype). c The neurons isolated from cerebral organoids were plated on coverslips and stained with Filipin III for cholesterol. Filipin III intensities were quantified in three fields of each coverslip and averaged (n = 4 coverslips/genotype), Scale bars, 20 μm. d Parental and APOE^−/− isogenic iPSCs were differentiated into radial glia/neural stem cells. Representative fluorescence microscopy images of the differentiated radial glia/neural stem cell stained with antibodies against Nestin and PAX6. Scale bar: 20 μm. e The mRNA levels of selective cholesterol biosynthesis genes in the radial glia/neural stem cell were quantified by RT-qPCR (n = 6 wells/genotype). f The radial glia/neural stem cells were plated on coverslips and stained with Filipin III for cholesterol. Filipin III intensities were quantified in three fields of each coverslip and averaged (n = 4 coverslips/genotype). Scale bars: 20 μm. Experiments were repeated in two independently differentiated batches. All data are expressed as mean ± SEM. Student’s t tests were performed to determine statistical significance. **p < 0.01, ***p < 0.001, n.s., not significant