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. Author manuscript; available in PMC: 2021 Jul 14.
Published in final edited form as: Nature. 2020 Jun 24;584(7819):98–101. doi: 10.1038/s41586-020-2428-0

Figure 2: pHi and glycolysis decrease during in vitro differentiation of human iPS-derived PSM cells.

Figure 2:

(a) Extra Cellular Acidification Rate (ECAR) and (b) Glycolytic ATP production rate in iPSCs and Day (D) 1-5 differentiated iPSCs cells. n=10.

(c) Snapshots from time-lapse imaging of eGFP-tagged β-catenin in differentiating iPSCs. (n=3 independent experiments). Scale bar = 50 μm.

(d) pHi analysis in human iPS cells differentiating to the PSM fate in vitro (n=3). One-way ANOVA followed by Tukey’s multiple comparisons test. **p < 0.01.

(e) Analysis of the pHi using BCECF dye in day 1-2 human iPS cells differentiated to the PSM fate in vitro and cultured for 3 h in CL culture medium at different pH (n=6 independent experiments). One-way ANOVA with Tukey’s multiple comparisons test. *p =0.023; ****p=7.26.e-6; ns p=0.999.

(f) pHi analysis in human iPS cells cultured for one day in CL medium followed by 24 h in CL medium with 2DG (n=4 for each condition). Unpaired two-tailed t-test: control vs 5mM 2DG, *P=0.01.

(g) SOX1 and AXIN2 expression in day 2 human PSM-like cells cultured for 3 h in CL medium at different pH (AXIN2 (Control, Acid, Alkali), n=5; SOX1 (Control, Acid); n=5, SOX1 (Alkali); n=6). Two-way ANOVA followed by Tukey’s multiple comparisons test. *p < 0.05; **p< 0.01.

(h-i) SOX1 and AXIN2 expression in human iPS cells cultured for one day in CL medium followed by a 24 h 2DG treatment (h) or in glucose-free CL medium (i) at normal or alkaline pH (n=3 experiments for each condition). Two-way ANOVA followed by Tukey’s multiple comparisons test. *p < 0.05; **p< 0.01.

(Control: pH7.0-7.2), Acid: pH6.3-6.5; Alkali: pH7.5-7.8) (a, d, f-h) Mean +/− SD. Exact p values can be found in the corresponding Source Data files.