Supplementary Figure 1: Screening strategy for the identification of endoderm subpopulations.
From: Generation of pancreatic β cells from CD177+ anterior definitive endoderm
(a) Screening work flow for the initial screen. (b-d) Representative FACS plots for CD177 and CD275 (b) labelling of differentiated day 4 DE cells with known endoderm markers (FOXA2 and CXCR4) revealed definitive endoderm (FOXA2+/CXCR4+) and mes-endoderm (FOXA2low/CXCR4-) subpopulations. (c-d) CD177 and CD275 expression profiles reveal different endoderm subpopulations (n = 1). (e) Immunofluorescent staining for CER1 with FOXA2 in DE cultures (n = 3, biologically independent experiments). Scale bar, 25 µm. (f) FACS analysis for CD275+/CER1+ and CD177+/CER1+ ADE cell populations at day 4 DE. (g-h) qPCR quantification for the mRNA expression of FOXA2 and SOX17 (g), CER1 and HHEX (h) in enriched CD177+ and CD275+ ADE subpopulations (ANOVA, (e-h) n = 3, biologically independent experiments). Data is represented as mean ±s.e.m; P<0.05 and P<0.01. Statistically non-significant results are not indicated in the figure.
