Fig. 6. Muscle-specific downregulation of SPoCk compromises CoQ biosynthesis, mitochondrial function and development in Drosophila.

a, b TXRF-based determination of Mn levels in third instar larvae (a) and adult female flies 1–3 days after eclosion (b). Values have been normalized to w¹¹¹⁸. n = 7 (larvae) and n = 4 (adults) per genotype (One n = 10 larvae or five flies). Means ± SEM. c, d Confocal micrographs (c) and corresponding quantification of fluorescence intensity (d) of w¹¹¹⁸ (mef2-Gal4 > ) and SPoCk-RNAi (mef2-Gal4 > UAS-SPoCk-RNAi) third instar larval muscle tissue stained with Mitotracker CMXRos (magenta) to determine mitochondrial transmembrane potential. Nuclei stained with DAPI (cyan); n = 10 (w¹¹¹⁸) or 9 (SPoCk-RNAi) animals. e, f Total CoQ content determined via HPLC-ECD analysis in third instar larval muscle tissue (e) and thorax muscle tissue (f) of w¹¹¹⁸ and SPoCk-RNAi flies 2 days after eclosion (not sex-sorted). Values have been normalized to CoQ₉ levels in w¹¹¹⁸; n = 9 (larval muscle) and n = 4 (adult thorax) per genotype (One n = 10 animals). Means ± SEM. g, h Developmental time of w¹¹¹⁸ and SPoCk-RNAi first instar larvae to adult reared on standard food or supplemented with CoQ₁₀ (g) or diHB (h), n = 208-274 animals (g), n = 232-335 animals (h). (i) Death of w¹¹¹⁸ and SPoCk-RNAi adults within 12 h after eclosion reared on standard food or supplemented with diHB, n > 13, with 24 flies per n. Box plots (d, e, i) show mean (x), median (line), first/third quartile (lower/upper bound), minimum/maximum within 1.5-fold IQR (lower/upper whisker), and outliers outside 1.5-fold IQR (circle/o). Details for statistical analysis (Supplementary Table 9) and source data are provided.