Fig. 1: Generation and characterization GBA-PD midbrain organoids.

a Schematic overview of the protocol used for the generation of midbrain organoids. mfNPCs, floorplate neural precursors. b GCase enzyme activity in differentiated MO cultures was significantly decreased (40% reduction) when compared with controls. The data represent a summary of three independent differentiation experiments per line each analyzed in triplicate at DIV30. Values are normalized to the average of controls per experiment. Wilcoxon T-test; ***p < 0.001. c GCase protein levels are not altered at DIV30. Representative western blot analysis and respective quantification. The data represent a summary of at least three independent differentiation experiments per line each analyzed in triplicate. Values are normalized to the average of controls per experiment. d Decreased percentage of colocalization of GCase with lysosomes in GBA-PD organoids at DIV30. GCase (red), LAMP1 (green). Representative confocal images with their respective zoomed region of interest (ROI) and their quantification (scale bar, 10 μm). The data represent a summary of three independent differentiation experiments for all cell lines, normalized to the average of controls per experiment. Wilcoxon T-test; *p < 0.05. e, f Representative western blot and quantification of expression of the autophagy markers p62 and LC3 at DIV30. Data represents a summary of at least five independent differentiation experiments. Values are normalized to the average of controls per experiment. Wilcoxon T-test; ***p < 0.001.