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. 2022 Nov 8;11:e82709. doi: 10.7554/eLife.82709

Figure 7. sloth1-sloth2 are important for mitochondrial function.

(A) Seahorse mitochondrial stress report for wildtype S2R+and dKO #1 cells. Error bars show mean with SD. N=6 for each genotype. (B) Quantification of basal OCR (timepoint 3) in panel A and including data from single KO and additional dKO cell lines. Significance of KO lines was calculated with a T-test compared to S2R+. Error bars show mean with SD. **** p≤0.0001. N=6 for each genotype. (C) Quantification of ATP levels in 3rd instar larvae. Error bars show mean with SEM. N=3 experiments. (D) Western blot from lysates of 3rd instar larval brains. (E–F) TEM images of sectioned adult photoreceptors (left) and quantification of mitochondria number (right). Mitochondria are indicated with red dots. Error bars show mean with SD. Sample size indicated on graph. (E) Adult flies are 4 weeks old and raised in a 12 hr light/dark cycle. (F) Adult flies are 3 days old and raised in a 12 hr light/dark cycle.

Figure 7.

Figure 7—figure supplement 1. Related to Figure 7.

Figure 7—figure supplement 1.

(A) Sequence analysis of single KO S2R+clones for sloth1 (clone 2F8) and sloth2 (clone 3A7). sgRNA and PAM site indicated by grey boxes. (B) PCR genotyping of four independently derived single cell dKO S2R+clones. (C–D) Seahorse mitochondrial stress test quantification of (C) ATP production and (D) Proton leak. Significance of KO lines was calculated with a T-test compared to S2R+. Error bars show mean with SD. ** p≤0.01, *** p≤0.001, **** p≤0.0001. N=6 for each genotype. (E) Confocal images of 3rd instar larval ventral nerve cord (VNC), axon bundles, and neuromuscular junction (NMJ). MN-Gal4 UAS-mitoGFP (MN>mitoGFP) (GFP) expresses mitochondrial-localized GFP in motor neurons. Neurons are stained with anti-HRP (magenta).
Figure 7—figure supplement 2. Related to Figure 7.

Figure 7—figure supplement 2.

(A–B) TEM images of sectioned adult photoreceptors. (A) Adult flies are 4 weeks old and raised on a 12 hr light/dark cycle. Mitochondria are indicated with red dots. (B) Adult flies are 3 days old and raised in a 12 hr light/dark cycle.