Fig. 2.

GM1 Ganglioside Storage and β-Gal Enzyme Activity in GLB1 knockout Cerebral Organoids.

(A) Representative images of isogenic control and GLB1 knockout cerebral organoids harvested at 5, 10, 15, and 20 weeks and stained with anti-GM1 ganglioside (green) and anti-β3 tubulin (red) antibodies. Nuclei were visualized with DAPI staining (blue). Arrows indicate areas of GM1 storage. Scale bars: white, 500 μm; yellow, 10 μm. (B) Quantification of GM1 fluorescent signal normalized to DAPI of GLB1 knockout organoids compared with isogenic control organoids. Data represent the mean relative GM1 expression ± SD, n ≥ 3 for both control and knockout organoids at each time point (^⁎⁎⁎p < .001, two-way Anova analysis between control and knockout values). (C) X-gal staining of isogenic control and GLB1 knockout cerebral organoids harvested at 15 weeks. Blue staining on the organoids indicates β-gal activity. Scale bars, 500 μm. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)