Fig. 3: Regulation of the segmentation clock by the NAD+/NADH ratio.
From: Metabolic regulation of species-specific developmental rates
a, ETC and relevant small molecule inhibitors. Adapted from BioRender.com (2021). Cyt c, cytochrome c. b, The HES7-Achilles oscillatory period in human PSM cells treated with DMSO control (n = 53), 20 nM rotenone (n = 23), 50 nM atpenin A5 (n = 36), 100 nM antimycin A (n = 26), 1 mM sodium azide (n = 30), 1 μM oligomycin (n = 44) or 1 μM FCCP (n = 55) for 24 h. Data are mean ± s.d. One-way ANOVA with Šidák correction. Rotenone, P = 1.1 × 10−8; atpenin, P = 6.4 × 10−14; antimycin, P = 1.85 × 10−23; azide, P = 1.88 × 10−42. c, Whole-cell NAD+/NADH ratio in human PSM cells treated with DMSO control, 20 nM rotenone, 100 nM antimycin A, 1 mM sodium azide and 1 mM sodium azide with 1 mM sodium pyruvate for 24 h. Data are mean ± s.d. n = 4 biological replicates. One-way ANOVA with Dunnett correction. Control versus azide, P = 1.1 × 10−5. d, The HES7-Achilles oscillatory period in human PSM cells treated with DMSO control (n = 67), 1 mM sodium azide alone (n = 46), azide with 1 mM sodium pyruvate (n = 27) or azide with 5 μM duroquinone (DQ) (n = 46). Data are mean ± s.d. One-way ANOVA with Tukey’s correction. Control versus azide, P = 7.4 × 10−14; control versus azide + pyruvate, P = 1.2 × 10−5; control versus azide + duroquinone, P = 7.5 × 10−14; azide versus azide + pyruvate, P = 1.4 × 10−13. e, NADH oxidation reaction catalysed by LbNOX28. f, Whole-cell NAD+/NADH ratio in human PSM cells transduced with a lentivirus expressing either mCherry alone or LbNOX with mCherry. Data are mean ± s.d. n = 8 biological replicates. Unpaired two-sided t-test. g, HES7-Achilles oscillatory period in human PSM cells transduced with a lentivirus expressing either mCherry alone (n = 113) or LbNOX with mCherry (n = 116). Data are mean ± s.d. Unpaired two-sided t-test: P = 3.7 × 10−10. h, MSGN1-Venus fluorescence during days 1–2 of human PSM differentiation, transduced with a lentivirus expressing either mCherry alone or LbNOX with mCherry. Data are mean ± s.e.m. n = 7 biological replicates. i, Cell cycle length in human PSM cells transduced with a lentivirus expressing either mCherry alone (n = 23) or LbNOX with mCherry (n = 25). Data are mean ± s.d. Unpaired two-sided t-test.
