Fig. 2: Mitochondrial abundance and morphologies are not altered in patient-derived cells.
From: A reversible state of hypometabolism in a human cellular model of sporadic Parkinson’s disease
a Quantification of total mitochondrial mass and functional mitochondria in hNPCs or b DAns. Total mitochondrial mass was visualized using immunostainings with an antibody against the ATP synthase (ATP5F1A). Functional mitochondria with an active membrane potential were visualized using a MitoTracker probe (200 nM for 20 min). Images are exemplarily shown for hNPCs of clone i1E4-R1-003 (Ctrl), and iR66-R1-007 (sPD); for DAns of clone i1E4-R1-003 (mitoTracker—Ctrl), iR66-R1-007 (mitoTracker—sPD), i1JF-R1-018 (ATP5F1A—Ctrl), and iJ2C-R1-015 (ATP5F1A—sPD). Scale bar = 20 μm. c Violin plots highlight some morphological characteristics of mitochondria in hNPCs or d DAns: number of mitochondria (count), mitochondrial area, skeleton length, number of branch points, and mean intensity. On average, 112 (hNPCs—ATP5F1A), 160 (hNPCs—mitoTracker), 370 (DAns—ATP5F1A), 360 (DAns—mitoTracker) cells per clone were analyzed. e To assess alterations in the mitochondrial fusion machinery, expression of the mitofusions MFN1, MFN2, and OPA1 was quantified in hNPCs by RT-qPCR. Cells were cultivated on the energy substrates used in the Seahorse XF analysis (25 mM glucose or 5 mM pyruvate). f To assess alterations in the mitochondrial fission machinery, expression of DRP1 and its phosphorylation on Ser616 was quantified in hNPCs by western blot. Protein levels were normalized to ACTB. g Western blots are exemplary shown for some hNPC clones. Boxplots display the median and range from the 25th to 75th percentile. Whiskers extend from the min to max value or to the most extreme data point which is no more than 1.5 times the interquartile range (c, d). Each dot represents one patient. n = 5 Ctrl and 7 sPD patient-derived cell clones, in triplicates. p-values were determined by linear mixed effects model (c, d); one-way ANOVA with Sidak’s Post hoc test (p-values are provided together with the source data) (e, f). *p < 0.05; **p < 0.01; ***p < 0.001. Source data are provided as a Source Data file.
