Fig. 6

FAC-sorted GP2⁺ cells give rise to ‘β-like’ cells in vitro. a Flow plots showing the GP2 profile at day 13 of the unsorted (presort) and fluorescence-activated cell sorted H1 cells, which were then cultured to generate β-like cells up to day 23. b qPCR analysis of GP2, NKX6-1, PDX1, and PTF1A. Expression levels normalized to TBP, and relative to adult pancreas (equal to 1, not shown). N = 4, error bars indicate s.e.m. *p < 0.05, **p < 0.01, ***p < 0.001, Student’s t-test. c Representative flow cytometry plots of day 23 cultures from H1-derived unsorted (presort), GP2⁺ or GP2⁻ populations stained with anti-NKX6-1 and anti-C-PEPTIDE (CPEP) antibodies. The bar graph shows the average percentage of NKX6-1⁺/C-PEPTIDE⁺ cells. N = 5, error bars indicate s.e.m. *p < 0.05, One-way ANOVA. d C-PEPTIDE (CPEP)/GLUCAGON (GCG)/TRYPSIN (TRYP) immunostaining of GP2⁺ and H1-derived unsorted (presort) cultures at day 23 of differentiation. Scale bar represents 50 μm. e Flow cytometry plots showing GP2 expression at day 13 and day 23 of differentiation in unsorted (presort) H1 cells. Abbreviations: GP2, pancreatic secretory granule membrane major glycoprotein 2; D13, day 13; NKX6-1, NK6 homeobox 1; PDX1, pancreatic and duodenal homeobox 1; PTF1A, pancreas specific transcription factor 1a; CPEP, c-peptide; GCG, glucagon; TRYP, trypsin; INS, insulin