Fig 1: Altered transcriptional profiles of apoE-deficient cerebral organoids implicating GBA and lipid-related pathways. RNA-seq was performed on parental control and isogenic APOE-/- iPSC-derived cerebral organoids at Day 90 (3 cerebral organoids were pooled and analyzed as one sample, n = 3 samples/each). a, Module-trait relationships between groups revealed by WGCNA are shown. The correlation coefficient (r) and the correlation p-value in the parentheses are indicated in each module. Orange indicates upregulation in APOE-/- organoids; blue indicates downregulation in APOE-/- organoids (upregulation in controls) b, Top gene ontologies enriched by the yellow module genes. c, Interaction of top 20 genes with the highest connectivity among each other in the yellow module. d–e, GBA mRNA expression (d) and ß-glucocerebrosidase (GCase) levels in the RIPA lysates (e) were quantified by RT-qPCR and Western blotting at Day 90, respectively. f, GCase activity in cerebral organoids was detected by GCase activity kit (Fluorometric). Data were normalized to protein concentrations. g, Amounts of LIMP1 in the iPSC-derived cerebral organoids at Day 90 were quantified by Western blotting. h, Top gene ontologies enriched by the green module genes. (i) Interaction of top 20 genes with the highest connectivity among each other in the green module. j, The lipid droplet accumulation was evaluated by BODIPY staining (Scale bar: 20 µm) with the fluorescent intensity quantified by Image J. All data are expressed as mean ± SEM (n = 4 organoids/each). k, Representative images of co-staining of BODIPY and Plin2 (lipid droplet membrane marker). Scale bar: 20 µm. l, Amounts of Plin2 in the iPSC-derived cerebral organoids at Day 90 were quantified by Western blotting. 3 cerebral organoids were pooled and analyzed as one sample. All data are expressed as mean ± SEM (n = 6 samples/each). Experiments were repeated in three independent differentiation batches. Mann–Whitney U tests were performed to determine statistical significance. *p < 0.05, **p < 0.01
Fig 2: Reduced GCase activity in monocytes from patients with Parkinson’s disease (PD). (a) Following sequencing, participants were split into wild type (blue circles, n = 80) or GBA1 missense mutation (red circles, n = 12) groups, and GCase activity compared by t-test. Data are mean ± SEM with dots indicating individual data points (b) Participants with GBA1 mutations were then removed from the analysis and the GCase activity in controls (blue circles, n = 41) and PD patients (red circles, n = 39) was again compared by t-test. Data are mean ± SEM with dots indicating individual data points (c) A univariate analysis was performed to determine if monocyte GCase activity was still reduced in the PD group when age and gender were included as covariates. Data are estimated marginal means ± SEM with the 95% confidence intervals indicated by dashed lines. (d) Receiver operator characteristic (ROC) curve indicating the sensitivity and specificity for monocyte GCase activity to classify PD.
Supplier Page from Abcam for Anti-GBA antibody