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. Author manuscript; available in PMC: 2020 Sep 11.
Published in final edited form as: Mol Cell. 2019 Feb 13;73(6):1127–1137.e5. doi: 10.1016/j.molcel.2019.01.013

Figure 2. PINK1 Accumulates on Mitochondria Enriched for the mt-ts Genome in the Ovary of Heteroplasmic Flies.

Figure 2.

(A) Confocal (left-two panels) and ratiometric images (right-two panels) of mid-stage egg chambers from mt-wt flies and heteroplasmic mt-ts (hetero.) flies expressing both PINK1-GFP and Tom20-mCherry at 29°C. Enlarged view of boxed areas shows PINK1-GFP colocalizing with Tom20-mCherry in mt-wt and heteroplasmic flies at 29°C. Ratiometric images (GFP/mCherry) show that PINK1-GFP level is elevated in heteroplasmic ovaries compared to mt-wt ovaries at 29°C. Enlarged view of boxed areas shows mitochondria (arrows) with much higher GFP/mCherry ratio than neighboring mitochondria in heteroplasmic flies at 29°C. Bars: 10 μm.

(B) Confocal imaging of two mitochondrial populations purified from pink1-GFP;; Tom20-mcherry (heteroplasmic) ovaries and sorted on the basis of GFP and mCherry intensity. Merged images are overlay of GFP and mCherry fluorescence with DIC channel. Note that GFP-high and GFP-low populations have similar level of Tom20-mCherry signal and are comparable in size. Bar: 1 μm.

(C) Mitochondria in two sorted populations have similar forward scatter (FSC-H) distribution, indicating they are comparable in size.

(D) Proportion of mt-ts genome in two sorted mitochondrial populations. High and low-GFP mitochondrial fractions from the same ovary are connected with a dotted line (n = 12). Note that the proportion of mt-ts genome is always higher in the high-GFP mitochondria fraction than the low-GFP fraction recovered from the same ovary.