Extended data Figure 5: Sodium Acetate treatment increases Wnt/β-catenin signaling in vivo and in vitro.

(a) Western blot analysis using anti-acetylated K49 β-catenin, anti-active β-catenin, anti-actin and anti-β-catenin antibodies. Whole cell extracts of day 2 hiPS 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 hiPS cells differentiated to PSM in vitro and treated with Sodium acetate (SA) in CL medium for 24 hours. Mean +/− SD (n=3). Two-way ANOVA followed by Tukey’s multiple comparisons test: MSGN1; Control versus 10mM 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 antibodies of whole cell extracts of 2-day chicken embryos cultured in chemically defined medium with 0 or 10mM Sodium acetate (SA) for 10 h. (n=3).

(d) Whole-mount in situ hybridization of 2-day chicken embryos cultured with 0 or 10 mM Sodium Acetate (SA) and hybridized with AXIN2 (control n=5, 10 mM Sodium acetate n=7). Scale bar: 100 μm.

(e) qPCR analysis of AXIN2, MSGN1, SOX2 and SAX1 expression in the posterior region of 2-day chicken embryos cultured with 0 or 10 mM Sodium Acetate. Data were normalized by control samples. Mean +/− SD (n=4). Two-sided unpaired t-test. AXIN2, p= 0.0070, MSGN1, p= 0.0298. *p<0.01, **p<0.001.

For gel source data, see Supplementary Figure 1