Supplementary Figure 6: H1 hESC pancreatic and endocrine differentiations of CD177⁺- and US-DE.

(a) Overview of differentiation protocol used to generate CD177-/US-β-cells. (b-e) Immunostainings for INS and NKX6.1 (b), GCG, INS and SST (c) and C-peptide and GLUT1 (d), INS, MAFA and NKX6.1 (e) in CD177- and US- β-cells ((b-e) n = 3, biologically independent experiments). Scale bars, 50 µm. (f) Representative flow cytometry contour plots of S4 and S7 cells generated from CD177⁺- and US-ADE/DE cells on H1 line and stained for indicated markers. (g,h) Percentage of cells expressing indicated markers (two-sided unpaired Student’s t-test, (g) n = 3, (h) n=13, n = 17 for INS+, 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.