Extended Data Fig. 5: Sodium acetate treatment increases WNT–β-catenin signalling in vivo and in vitro.
From: Intracellular pH controls WNT downstream of glycolysis in amniote embryos
a, Western blot analysis using anti-acetylated K49 β-catenin, anti-active β-catenin, anti-actin and anti-β-catenin. Whole-cell extracts of day 2 human iPS cells differentiated to PSM in vitro in CL medium and treated with sodium acetate (SA) for 24 h (n = 3). b, qPCR analysis of SOX1 and MSGN1 mRNA expression in day 2 human iPS cells differentiated to PSM in vitro and treated with SA in CL medium for 24 h. Mean ± s.d. is shown (n = 3). Two-way ANOVA followed by Tukey’s multiple comparisons test. MSGN1 control versus 10 mM SA: P = 0.003. **P < 0.01. c, 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 10 mM SA for 10 h (n = 3). d, Whole-mount in situ hybridization of 2-day-old chicken embryos cultured with 0 or 10 mM SA and hybridized with AXIN2 (control: n = 5 and 10 mM SA: n = 7). Scale bars, 100 μm. e, qPCR analysis of AXIN2, MSGN1, SOX2 and SAX1 expression in the posterior region of 2-day-old chicken embryos cultured with 0 or 10 mM SA. Data were normalized by control samples. Mean ± s.d. is shown (n = 4). Two-sided, unpaired t-test. AXIN2: P = 0.0070 and MSGN1: P = 0.0298. *P < 0.01 and **P < 0.001. For gel source data, see Supplementary Fig. 1.
