Fig. 3

Cisd overexpression blocks mitophagy flux. A Immunoblot analysis of whole fly lysates from 2-day-old flies of the indicated genotypes, analysed for pUb, and Tubulin or total protein levels as loading controls. Blot is representative of 3 replicate experiments. Cisd overexpression was driven by da-GAL4. B Confocal microscopy analysis of adult Drosophila flight muscle from 2-day-old flies of WT control, parkin mutant and Cisd overexpression driven by da-GAL4 immunostained for mitochondria (ATP5A) and pUb. C–F Confocal analysis of mitophagy reporter mito-QC (OMM-localised tandem RFP-GFP) of WT or Cisd overexpression driven by nSyb-GAL4, in larval (C, D) or adult (E, F) neurons with ‘red-only’ mitolysosomes shown. D, F Number of mitolysosomes quantified shown in C and E. Data points indicate individual animals analysed. Statistical analysis: unpaired t-test; **P < 0.01; ****P < 0.0001. Scale bars = 10 μm