Extended Data Fig. 4: Kinetics of WNT–β-catenin signalling during human iPS cell differentiation to the PSM.
From: Intracellular pH controls WNT downstream of glycolysis in amniote embryos
a, Immunohistochemistry showing the dynamic expression of β-catenin and Venus (YFP) proteins in human MSGN1-Venus iPS reporter cells differentiated to the PSM fate in vitro (n = 3). Hoechst labelling of the nuclei is shown in blue. Scale bars, 30 μm. b, Quantification of the intensity of nuclear localization of β-catenin shown in a using Fiji. Mean ± s.d. is shown (D0, D1 and D4: n = 3; D2 and D3: n = 4). One-way ANOVA followed by Tukey’s multiple comparisons test: D1 versus D2: P = 0.0411, D2 versus D3: P = 0.0038, D2 versus D4: P = 0.0009. *P < 0.05, **P < 0.01 and ***P < 0.001. c–e, qPCR analysis comparing the expression level of MSGN1 (c), TBX6 (d) and AXIN2 (e) of human iPS cells differentiating to the PSM fate in vitro. Values were normalized by the results of differentiation at D0. Mean ± s.d. is shown (n = 3). One-way ANOVA followed by Tukey’s multiple comparisons test: TBX6 D0 versus D2: P < 0.0001, D0 versus D3: P < 0.0001, D3 versus D4: P < 0.0001, D4 versus D10: P = 0.0466; AXIN2 D0 versus D1: P = 0.0006, D0 versus D3: P < 0.0001, D3 versus D4: P = 0.0003, D4 versus D10: P = 0.0318 *P < 0.05, ***P < 0.001 and ****P < 0.0001.
