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. Author manuscript; available in PMC: 2021 Jul 15.
Published in final edited form as: Free Radic Biol Med. 2020 Feb 1;152:797–809. doi: 10.1016/j.freeradbiomed.2020.01.177

Figure 3. Mitochondrial dysfunction in pmp-4(ok396) animals.

Figure 3

WT and pmp-4(ok396) worms were incubated at the L4 larval stage with (A) the mitochondrial complex I inhibitor Paraquat (PQT) (n=51 to 54 animals by condition), the mitochondrial complex II inhibitors (B) 3-nitropropionic acid (3-NP) (n=15 to 20 animals by condition), (C) thenoyltrifluoroacetone (TTFA) (n=18 to 20 animals by condition), (D) the complex III inhibitor (antimycin A) (n=20 animals by condition) and (E) the complex IV inhibitor (sodium azide)(n=20 animals by condition). The lethality of the worms after the treatment was evaluated as described in the methodology. (F-Q). At L4, WT and pmp-4(ok396) nematodes were treated with CoQ (1 mg/ml) or MitoQ (5 μg/ml) for 7 days. Total ROS levels were measured by quantifying the fluorescence emission of the H2DCFDA probes in living animals in (F) WT (n=42), (G) pmp-4(ok396)(n=35), (H) WT + CoQ (n=32), (I) pmp-4(ok396) + CoQ (n=30), (J) WT + MitoQ (n=47), and (K) pmp-4(ok396) + MitoQ (n=32) animals at L4+7 days. Values are normalized to the untreated WT nematodes. Scale = 50 μm. Bright field images of the posterior part of the worm stained with Sudan Black in (L) WT (n=46), (M) pmp-4(ok396)(n=61), (N) WT + CoQ (n=60), (O) pmp-4(ok396) + CoQ (n=60), (P) WT + MitoQ (n=30), and (Q) pmp-4(ok396) + MitoQ (n=27) worms at L4+7 days. Lipids are evident as black droplets, labelled by a black arrow. Scale = 10 μm. Semi-quantitative analysis of the lipid droplets with a diameter >= 5 μm under the indicated conditions. Data represent the mean ± SD. Statistical analysis was carried out with Student’s t-test (*P<0.05; **P<0.01; ***P<0.001) for A-E. Statistical analysis was carried out with two-way ANOVA, followed by Tukey’s post hoc test (*P<0.05; **P<0.01; ***P<0.001) for F-Q.