Extended Data Fig. 7: Regulation of blastocyst size by cortical tension, tight-junction permeability, ion influx and cell–cell adhesion.
From: Hydraulic control of mammalian embryo size and cell fate
a, Dose-dependent volume expansion rate in Bb(−)-treated embryos (red, n = 15, 8 and 4 embryos for 5, 12.5 and 25 μM, respectively) and m−/− Myh9+/− embryos (blue, n = 12 embryos), compared to the wild type (black, n = 23 embryos). b, c, Maximum cavity diameter reached for wild type (black, n = 14 embryos), m−/− Myh9+/− (blue, n = 12 embryos) and embryos treated with Bb(−) (red) in various concentrations (n = 15, 8 and 4 embryos for 5, 12.5 and 25 μM, respectively) (b). These embryos all have similar numbers of cells (c), which shows that the reduced cavity size is not due to a reduced number of cells. n = 28, 24 and 31 embryos for wild-type, m−/− Myh9+/− and Bb(−)-treated embryos, respectively. d, Volume expansion rate in CPE- (red, n = 12 embryos), ouabain- (blue, n = 17 embryos), hypertonic- (pink, n = 28 embryos) and ECCD1-treated (green, n = 23 embryos) embryos compared to controls (black, n = 23 embryos). e, Maximum cavity diameter for CPE- (red, n = 14 embryos), ouabain- (blue, n = 17 embryos), hypertonic- (pink, n = 25 embryos) and ECCD1-treated (green, n = 16 embryos) embryos compared to controls (black, n = 14 embryos). f, ECCD1-treated embryos have similar numbers of cells to those of controls (dark blue, E5.0), whereas CPE-treated embryos have slightly higher numbers of cells than do control embryos (black, E4.25), despite their reduced cavity size. n = 17 and 26 embryos for 313 (CPE131, used as control) and CPE-treated embryos; n = 21, 26 and 28 embryos for control, ouabain- and hypertonic-treated embryos; and n = 18 and 17 embryos for control and ECCD1-treated embryos, respectively. g–i, Embryos treated with Y-27632 (a Rho-kinase inhibitor that targets actomyosin contractility) showed a reduced expansion rate (g) and cavity size (h), despite having cell numbers that were similar to those of controls (i). n = 14, 13, 11 and 10 embryos for controls, 10, 20 and 40 μM Y-27632-treated embryos, respectively, in g and h; n = 19 and 15 for control and Y-27632-treated embryos, respectively, in i. All box plots show median line, 25th and 75th percentiles, and whiskers extending to maximum and minimum data points. Mann–Whitney U test. ****P < 1 × 10−5.
