Fig. 4: PITRM1−/− cerebral organoids display the main pathological features of AD pathology and the induction of the mitochondrial stress response. | Molecular Psychiatry

Fig. 4: PITRM1−/− cerebral organoids display the main pathological features of AD pathology and the induction of the mitochondrial stress response.

From: Loss of function of the mitochondrial peptidase PITRM1 induces proteotoxic stress and Alzheimer’s disease-like pathology in human cerebral organoids

Fig. 4

a Western blot of APP in 2-month-old PITRM1+/+ and PITRM1−/− cerebral organoids. b Quantification of APP protein levels in cerebral organoids (mean + SEM; *p < 0.05, two-tailed t test, n = 5). c Quantification of Aβ species in the supernatant of PITRM1+/+ and PITRM1−/− iPSC-derived cerebral organoids, as performed by Meso Scale immunoassay (mean + SEM; *p < 0.05, **p < 0.01, ***p < 0.0001, two-tailed t test, n = 7). d Representative western blots of phospho-tau (Thr231, Ser202 and Thr205, Thr181 phosphorylation sites) and total tau in PITRM1+/+ and PITRM1−/− cerebral organoids; total tau and β-Actin were used as the loading controls. e Quantification of phospho-tau protein levels in cerebral organoids relative to the loading control total tau/β-Actin (mean + SEM; *p < 0.05, two-tailed t test, n = 3–5). f Immunostaining for β-TUBIII (green) and cleaved caspase-3 (cCASP3, red) in PITRM1+/+ and PITRM1−/− 2-month-old cerebral organoids. Cell nuclei were counterstained with DAPI (blue). Scale bar, 100 µm. g Analysis of the ratio of cCASP3-positive cells relative to the total number of β-TUBIII/DAPI cells in 2-month-old cerebral organoids (mean + SEM; ***p < 0.001, two-tailed t test, n = 3). h Phalloidin (red) and BTA-1 (blue) staining in cerebral organoids. Representative confocal images are shown. Scale bars, 100 μm. i Quantification of the size of BTA-1-positive areas in cerebral organoids (mean + SEM; **p < 0.01, two-tailed t test, n = 3). j MAP2 (green) and ubiquitin (red) immunostaining in cerebral organoids. Representative confocal images are shown. Cell nuclei were counterstained with DAPI (blue). Scale bars, 100 μm. k Quantification of ubiquitin mean fluorescent intensity (mean + SEM; *p < 0.05, two-tailed t test, n = 3). l Gene expression levels of mitochondrial stress response genes in PITRM1+/+ and PITRM1−/− cerebral organoids (mean + SEM; *p < 0.05, **p < 0.01, ***p < 0.001, two-tailed t test, n = 4–6).

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