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. 2014 May 22;23(19):5227–5242. doi: 10.1093/hmg/ddu244

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Figure 8. — CG8334 knockdown rescues parkin RNAi phenotypes in Drosophila. (A–D) Morphological analysis of mitoGFP-expressing indirect flight muscle from 1-week-old control (w¹¹¹⁸; UAS-mitoGFP/+; mef-2-GAL4/+), CG8334 RNAi (w¹¹¹⁸; UAS-mitoGFP/+; CG8334 RNAi/mef-2-GAL4), parkin RNAi (w¹¹¹⁸; UAS-mitoGFP/parkin RNAi; mef-2-GAL4/+) and (parkin + CG8334) RNAi (w¹¹¹⁸; UAS-mitoGFP/parkin RNAi; mef-2-GAL4/CG8334 RNAi) flies. (A) Confocal images of mitoGFP in indirect flight muscle. Scale bar, 10 µm. (B) Quantification of the number of mitochondrial clumps larger than 5 µm² in mitoGFP-expressing indirect flight muscle, normalized to control (n = 6 flies per genotype). *P < 0.05 compared with Control; ^#P < 0.05 compared with CG8334 RNAi; ^§P < 0.05 compared with (parkin + CG8334) RNAi. (C) Quantification of mitochondrial clump size in mitoGFP-expressing muscle (n = 6 flies per genotype). *P < 0.001 compared with Control; ^#P < 0.001, ^&P < 0.05 compared with CG8334 RNAi; ^§P < 0.01 compared with (parkin + CG8334) RNAi. (D) EM images of indirect flight muscle. The bottom row shows magnifications of the boxed areas in the row above. Scale bars, first row: 5 µm; second row: 1 µm. (E and F) Mitochondrial membrane potential (Δψm) at third instar larval boutons of control (w¹¹¹⁸; tub-GAL4/+), CG8334 RNAi (w¹¹¹⁸; ; CG8334 RNAi/tub-GAL4), parkin RNAi (w¹¹¹⁸; parkin RNAi/+; tub-GAL4/+) and (parkin + CG8334) RNAi (w¹¹¹⁸; parkin RNAi/+; CG8334 RNAi/tub-GAL4) flies was imaged using JC-1, a potentiometric green fluorescent dye that shifts to red fluorescence within mitochondria with a normal negative Δψm (red, JC-1 aggregates; green, JC-1 monomers). Scale bar, 5 µm. (F) Quantification of red over green fluorescence intensity in synaptic mitochondria normalized to control (n ≥ 15 synapses per genotype). *P < 0.005 compared with control. (G) Analysis of negative geotaxis in 2-week-old flies with the same genotypes as in (E) and (F) (n = 10 batches of 10 flies per genotype). The percentage flies that cross a 4 cm line in 15 s after tapping the flies down is indicated. *P < 0.05 compared with Control; ^#P < 0.01 compared with CG8334 RNAi; ^§P < 0.005 compared with parkin + CG8334 RNAi.