Extended Data Fig. 3: Inhibiting lactate transporters downregulates WNT signalling.
From: Intracellular pH controls WNT downstream of glycolysis in amniote embryos
a, Whole-mount in situ hybridization of a 2-day-old chicken embryo hybridized with MCT1 (n = 4). Scale bar, 100 μm. b, Comparison of lactate amounts in cellular extracts of the posterior region of 2-day-old chicken embryos cultured for 10 h in chemically defined medium with or without 5 mM CNCn (n = 3). Mean ± s.d. is shown. Two-sided, unpaired t-test, P = 0.0292. *P < 0.05. c, Whole-mount in situ hybridization of 2-day-old chicken embryos cultured with 0 mM (control (CTL)) or 5 mM CNCn and hybridized with AXIN2 (control: n = 8, 5 mM CNCn: n = 7). Scale bars, 100 μm. d, qPCR analysis of MSGN1, SAX1, SOX2, T and AXIN2 expression in the posterior region of 2-day-old chicken embryos cultured with or without 5 mM CNCn (n = 3 for each gene). Data were normalized by control samples. Mean ± s.d. is shown. Two-sided, unpaired t-test. AXIN2: P = 0.0197, T: P = 0.0270, SOX2: P = 0.0458. *P < 0.05. e, Comparison of AXIN2 and SOX1 mRNA expression in day 2 human iPS cells differentiated in vitro and cultured for 24 h in CL medium containing 5 mM CNCn or vehicle control (DMSO). n = 3 biological replicates. Mean ± s.d. is shown. Two-way ANOVA followed by Tukey’s multiple comparisons test: ***P = 0.0004 and **P = 0.0067. n = 3. f, Western blot analysis using anti-acetylated K49 β-catenin, anti-active β-catenin, anti-actin and anti-β-catenin of whole-cell extracts of 2-day-old chicken embryos cultured in chemically defined medium with 0 or 5 mM CNCn for 10 h (n = 3). For gel source data, see Supplementary Fig. 1.
