FIGURE

Fig. 6

ID

ZDB-FIG-200306-90

Publication

Luciani et al., 2020 - Impaired mitophagy links mitochondrial disease to epithelial stress in methylmalonyl-CoA mutase deficiency

Other Figures

All Figure Page

Back to All Figure Page

Fig. 6

Rescue of mitochondrial function and homeostasis by re-expressing PINK1 in MMA kidney cells.

a–c Cells were transduced with adenovirus particles expressing mitochondrially targeted GFP (Ad-mito-GFP, green) and with adenovirus particles bearing either Null or HA-PINK1. Cells were immunostained for HA (red) and Parkin (red). a Representative images and b quantification of numbers of Parkin⁺ structures in a cell. Number of control cells transduced with Null (n = 25) or HA-PINK1 (n = 39) and MMA cells transduced with Null (n = 57) or HA-PINK1 (n = 55). c Quantification of mito-GFP/Parkin⁺ structures (expressed as the percentage of total mitochondria); n ≥ 4 randomly selected and non-overlapping fields of views per each condition. Nuclei counterstained with DAPI (blue). d, e Null and HA-PINK1-expressing cells were transduced with adenovirus particles bearing mt-Keima. d Confocal microscopy-based quantification of the red/green fluorescence intensity ratio in a cell. Number of control cells transduced with Null (n = 46) or HA-PINK1 (n = 38) and MMA cells transduced with Null (n = 51) or HA−PINK1 (n = 54). e Representative immunoblotting and quantification of the indicated mitochondrial proteins; n = 5 biologically independent experiments. f, g Cells were loaded with (f) TMRM (green) or (g) MitoSOX (green) and MitoTracker (red), and analysed by confocal microscopy. Quantification of TMRM/MitoTracker or MitoSOX/MitoTracker fluorescence intensity ratio in a cell. Number of control cells transduced with Null (n = 43) or HA-PINK1 (n = 62) and MMA cells transduced with Null (n = 83) or HA-PINK1 (n = 84) for TMRM/MitoTracker. Number of control cells transduced with Null (n = 38) or HA-PINK1 (n = 56) and MMA cells transduced with Null (n = 59) or HA-PINK1 (n = 64) for MitoSOX/MitoTracker. h Oxygen consumption rate (OCR) and individual parameters for basal respiration, ATP production and maximal respiration. OCRs were measured at baseline and after the sequential addition of Oligomycin (Oligo, 1 μM), FCCP (0.5 μM) and Rotenone (Rot; 1 μM) + Antimycin A (Ant; 1 μM). Values in a–h are pooled from three biologically independent experiments. Plots represent mean ± SEM. One-way ANOVA followed by Bonferroni’s post hoc test, *P < 0.05 and ***P < 0.001 relative to control or MMA cells transduced with Null in b, d, f and g. Two-tailed Student’s t-test, *P < 0.05, **P < 0.01, ***P < 0.001 and ^#P < 0.0001 relative to control and MMA cells transduced with Null in c, e and h. Scale bars,10 μm. NS non-significant. Source data are provided as a Source Data file.

Expression Data

Expression Detail

Antibody Labeling

Phenotype Data

Phenotype Detail

Acknowledgments

This image is the copyrighted work of the attributed author or publisher, and ZFIN has permission only to display this image to its users. Additional permissions should be obtained from the applicable author or publisher of the image. Full text @ Nat. Commun.