Fig. 3: Impaired mitochondrial respiration and altered morphology in ABCA7−/− iPSC-derived neurons.
From: ABCA7 deficiency causes neuronal dysregulation by altering mitochondrial lipid metabolism
A Immunocytochemical staining of iPSC-derived neurons 6 weeks after differentiation from NPCs for neuron markers with βIII-tubulin and MAP2. Nuclei were stained with DAPI. Scale bar: 100 µm. B, C Mitochondrial respiration in neurons derived from isogenic control and ABCA7−/− iPSCs (#1 and #2) were measured by Mito Stress Test Kit through Seahorse XFe96 Extracellular Flux Analyzer 6 weeks after differentiation. The OCR measurements were normalized to cell density determined by nuclear DAPI staining in each well (n = 5 technical replicates/line). A.U., arbitrary unit. D, E The iPSC-derived neurons were stained for MitoSOX and MitoTracker. Mitochondrial ROS was assessed as relative MitoSOX/MitoTracker ratio (n = 4–6 technical replicates/line). Scale bars: 200 nm. F, G Representative electron microscope images of mitochondria in the iPSC-derived neurons are shown. The length, width, area, perimeter, and circularity of mitochondria in neurons derived from isogenic control and ABCA7−/− iPSCs (#1) were measured with ImageJ. (Control: n = 134 mitochondria from 3 clones, ABCA7−/−: n = 107 mitochondria from 3 technical replicates). Scale bar: 100 nm. Data represents mean ± SEM. *p < 0.05, **p < 0.01 by two-tailed student’s t-test.
