Extended Data Fig. 1: The segmentation clock-period is cell-autonomous even in chimeric conditions.
From: Metabolic regulation of species-specific developmental rates
a. Representative micrographs of MSGN1-Venus fluorescence in mouse (left) and human (right) PSC-derived PSM cells on day 2 of differentiation. Note the reporter is cytoplasmic in mouse cells but nuclear in human cells. Similar results were obtained n = 15 times. Scale bar = 400 μm. b. Period of segmentation clock oscillations in primary PSM tissue dissected from E9.5 mouse embryos carrying the LuVeLu reporter (n = 18) and PSC-derived PSM expressing the Hes7-Achilles reporter (n = 24). Mean ±SD. Unpaired two-sided t-test. c. Cell cycle duration in primary PSM tissue dissected from E9.5 mouse embryos (n = 27) and PSC-derived PSM (n = 33). Mean ±SD. Unpaired two-sided t-test. d. Instantaneous HES7-Achilles oscillatory period over the course of 16 h as calculated by Hilbert transformation for mouse PSM cells co-cultured with mouse (n = 10) or human (n = 7) non-reporter PSM cells. Mean ±SEM. e. Instantaneous HES7-Achilles oscillatory period over the course of 16 h as calculated by Hilbert transformation for human PSM cells co-cultured with human (n = 11) or mouse (n = 8) non-reporter PSM cells. Mean ±SEM.f. Mean segmentation clock period as calculated by Hilbert transformation for mouse (left) or human (right) PSM cells co-cultured with either mouse or human non-reporter PSM cells. Mean ±SD. n = 10 (mouse-mouse), n = 9 (mouse-human), n = 8 (human-mouse), n = 10 (human-human). One-way ANOVA with Šidák correction. g. Mean amplitude of HES7-Achilles oscillations in mouse (left) or human (right) PSM cells co-cultured with either mouse or human non-reporter PSM cells. Mean ±SD. n = 10 (mouse-mouse), n = 9 (mouse-human), n = 8 (human-mouse), n = 10 (human-human). Kruskal-Wallis test with Dunn’s correction. h. Representative single-cell tracks of HES7-Achilles fluorescence for mouse PSM cells cultured with non-reporter mouse PSM cells. i. Representative single-cell tracks of HES7-Achilles fluorescence for mouse PSM cells cultured with non-reporter human PSM cells. j. Representative single-cell tracks of HES7-Achilles fluorescence for human PSM cells cultured with non-reporter human PSM cells. k. Representative single-cell tracks of HES7-Achilles fluorescence for human PSM cells cultured with non-reporter mouse PSM cells.
