Fig. 4
JNCL cerebral organoids recapitulate disease hallmarks in the neuronal tissue. a Representative TEM pictures highlighting increased presence of autophagic vacuoles (AVs, dashed yellow line) in the CLN3Q352X organoids. Scale bars, 1 μm. b Quantification of AV area, measured on ImageJ by region of interest (ROI) definition, reveals a significant increase in the size of the vacuoles in the CLN3Q352X organoids. Comparison from 14 Control and 20 CLN3Q352X random AV profiles was done using Mann-Whitney test, *p < 0.05. Data points indicate single AVs measured per condition. c Representative TEM pictures of fingerprint inclusions (FPPs) detected exclusively in the CLN3Q352X organoids. Scale bar, 500 nm. d Representative EM pictures of curvilinear bodies (CVB) found exclusively in the CLN3Q352X organoids. Scale bar, 500 nm. e Representative western blot staining for TPP1 showing a slight increase in the CLN3Q352X organoids. f Quantification of the TPP1 levels from western blots. Data points represent individual organoids (n = 12 organoids from at least three independent derivations) (Welch’s test; **p < 0.01). g ELISA quantification of CTSD protein levels. Data points represent single different organoids (n = 8 organoids generated in at least three independent derivations) (Welch’s test; **p < 0.01). h Astrocytosis shown by increased GFAP positive (green) staining in the CLN3Q352X organoids compared to the controls. Scale bars, 100 μm. i Image analysis of GFAP and MAP 2 positive staining in organoid sections, normalized by nuclear (Hoechst positive) staining. Data points represent different organoid sections (n = 8 Control and 9 CLN3Q352X sections taken from different organoids generated in at least three independent derivations). Comparison evaluated by Kolmogorov-Smirnov test (*p < 0.05; ns, not significant). Data in B, F, G and I is shown as mean ± SEM