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. 2019 Nov 21;10:5284. doi: 10.1038/s41467-019-13172-8

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Fig. 5 — NAD⁺ ameliorates premature aging in WS through DCT-1 and ULK1-dependent mitophagy. a Mitochondrial content was evaluated by quantifying MitoTracker green pixels of stained N2 and wrn-1(gk99) worms at adult D1 and D7 (n = 80 worms; Two-way ANOVA). values pooled from three independent biological repeats. Data from Vehicle-treated worms are also used in Fig. 1h. b Quantified scores of muscle mitochondrial morphology of adult D7 N2 and wrn-1(gk99) worms. A myo-3::gfp reporter gene was expressed in body wall muscles. (n = 20 worms/group; Two-way ANOVA). c, d Relative mitophagy rate in adult D7 N2 and wrn-1(gk99) worms. Mitophagy events were calculated as the co-localization between DsRed::LGG-1 and DCT-1::GFP in muscle cells. (n = 20 worms for each condition; Two-way ANOVA). e Effects of NR or UA treatment on pharyngeal pumping in N2 and the wrn-1(gk99) worms at adult D4 and D6. (n = 3 biologically independent experiments with 10–20 worms for each condition; Two-way ANOVA). f Pharyngeal pumping rates in adult D7 worms of designated groups. (n = 20 worms/group; Two-way ANOVA). g, h mRNA levels of dct-1 (d) and unc-51 (e) in adult D7 worms. (n = 3 biologically independent experiments; Two-way ANOVA). i Flow cytometry quantification of relative mitophagy incidence in HT01, WS01, and WRN-KD cells under different conditions. For siRNA control, siRNA-vector was added cells. (n = 3 biologically independent experiments; Two-way ANOVA). j Western blot data showing changes of expression of designated proteins. Source data are provided as a Source Data file. Two-way ANOVA followed by Tukey’s post-hoc tests. Data are shown in mean ± S.E.M. *p < 0.05, **p < 0.01, ***p < 0.001.