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. 2020 Nov 2;11:5540. doi: 10.1038/s41467-020-19264-0

Fig. 7. Isogenic conversion of APOE4 to APOE3 attenuates AD-related phenotypes in cerebral organoids.

Fig. 7

The iPSC-derived cerebral organoids from an AD patient carrying APOE ε4/ε4 (Par-E4/4) and the APOE ε3/ε3 isogenic line (Iso-E3/3) were analyzed at week 12. a The perimeters of cerebral organoids were measured (n = 10). b Representative images of cellular apoptosis with the immunostaining for cleaved CASP3. Scale bar: 100 μm. c Cleaved CASP3 and CASP3 levels in the lysates were analyzed by western blotting and quantified (p = 0.0043). d–f Amounts of Aβ40 (d: p = 0.0043) and Aβ42 (e: p = 0.0152) in RIPA fraction were measured by ELISA. Data were normalized to individual protein concentrations. The ratio of Aβ42/Aβ40 (f) was calculated accordingly. g, h Amounts of apoE in RIPA (g) and FA (h: p = 0.0411) were measured by ELISA. Data were normalized to individual total protein concentrations. i Representative images of the immunostaining for p-tau with AT8 antibody. Scale bar: 100 µm. j Total tau and p-tau in RIPA lysates were analyzed by western blotting and quantified. The p-tau detected by AT8 antibody and PHF1 antibody were normalized to total tau (AT8: p = 0.0152, PHF1: p = 0.0411). k, l Amounts of p-tau in RIPA (k: p = 0.0152) and FA (l: p = 0.0022) were measured by ELISA. Data were normalized to individual total protein concentration. m G3BP, levels in RIPA lysates were analyzed by western blotting and quantified. The levels of G3BP were normalized to β-actin (p = 0.0022). c–h, j–m Lysates of 4–5 cerebral organoids were analyzed as one sample. All data are expressed as mean ± SEM (n = 6). Two-sided Mann–Whitney U tests were performed to determined statistical significance, *p < 0.05, **p < 0.01.