Fig. 7

Increased proportion of small-volume endosomes in BIN1 null mutant hiNs. a– b’’ Representative images of BIN1 WT and KO hiNs in 6-week-old 2D cultures immunolabeled with antibodies against MAP2 (red, a and b) and EEA1 (green, a’ and b’) and stained with Hoechst 33258 (blue, a’’ and b’’) (Scale Bar = 10 µm). c Plot showing the quantification of the number of EEA1⁺ puncta per 1000 µm³ of MAP2⁺ surface (N = 3 independent cell cultures; BIN1 WT hiNs: 21.15 ± 9.94 EEA1⁺ puncta per 1000µm³ of MAP2⁺ surface; BIN1 KO hiNs: 25.31 ± 16.58 EEA1⁺ puncta per 1000µm³ of MAP2⁺ surface; p = 0.84, Unpaired t-test). d Plot showing the cumulative distribution of EEA1⁺ puncta volumes in BIN1 WT (blue line) and KO (red line) hiNs (N = 3 independent cell cultures; Kolmogorov–Smirnov test, ****p < 0.0001). e–f’’ Coronal sections of 190-days-old BIN1 WT and KO cerebral organoids immunolabeled with antibodies against MAP2 (red, e and f) and EEA1 (green, e’ and f’) and stained with Hoechst 33258 (blue, e’’ and f’’). g Quantification of the number of EEA1⁺ puncta per 1000 µm³ of MAP2⁺ surface (N = 3 organoids per genotype; cerebral organoids: BIN1 WT hiNs: 66.72 ± 18.13 EEA1⁺ puncta per 1000µm³ of MAP2⁺ surface; BIN1 KO hiNs: 73.18 ± 12.47 EEA1⁺ puncta per 1000µm³ of MAP2⁺ surface; Unpaired t-test, p = 0.96). h Cumulative distribution of EEA1⁺ puncta volumes in BIN1 WT (blue line) and KO (red line) hiNs (N = 3 organoids per genotype; Kolmogorov–Smirnov test, ***p < 0.001)