Fig. 5

APOE4 induces loss-of-function phenotypes in neurogenesis and cholesterol metabolism in the iPSC-derived cerebral organoids. The iPSC-derived cerebral organoids from an AD patient carrying APOE ε4/ε4 and the APOE ε3/ε3 isogenic line were analyzed at Day 90. a The mRNA levels of cerebral layer markers (DCX, SLC17A7, TBR1, BCL11B and SATB2) and astrocytic markers (S100B and GFAP) were quantified by RT-qPCR. Three cerebral organoids were pooled and analyzed as one sample (n = 5–8 samples/genotype). b Phosphorylation levels of eIF2α in APOE3 and APOE4 cerebral organoids at Day 90 were quantified by Western blotting. Three cerebral organoids were pooled and analyzed as one sample (n = 3 samples/genotype). c, d The mRNA levels of selective cholesterol biosynthesis genes in the neurons (c) and astrocytes (d) sorted from cerebral organoids were quantified by RT-qPCR (n = 6 wells/genotype). e, f The isolated neurons (e) and astrocytes (f) were plated on coverslips and stained with Filipin III for cholesterol. Filipin III intensities were quantified in 5 fields of each coverslip and averaged (n = 5 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 < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. For raw data see Additional file 1: Fig. S11