Fig. 2

Validation of the PP marker GP2 by flow cytometry qPCR and immunocytochemistry. a,b Flow cytometry analyses of undifferentiated hESCs, and day 13 PP and PH cultures. Cells were stained with anti-GP2. N = 11 for hESC, N = 9 for PP and N = 8 for PH, error bars indicate s.e.m. ***p < 0.001,Student’s t-test. c qPCR analyses of GP2 in undifferentiated hESCs, and day 13 PP and PH cultures. Expression levels normalized to TBP, and relative to adult pancreas (equal to 1, not shown). N = 3 for hESC and N = 4 for PP and PH, error bars indicate s.e.m. *p < 0.05, **p < 0.01, Student’s t-test. d GP2 immunostaining of undifferentiated hESCs, and day 13 PP and PH cultures. Scale bar represents 50 μM. e,f NKX6-1/GP2/INSULIN (INS) immunostaining of human pancreata at gestational weeks 33, 37, and 39.5, scale bar represents 100 μm. White box in top panel indicates area enlarged in bottom panel, scale bar represents 50 μm. g PTF1A/GP2/NKX6-1 immunostaining of human pancreata at gestational week 37. White dotted line indicates branching of the developing pancreas. Yellow arrow heads indicate PTF1A⁺/GP2⁺/NKX6-1⁺ cells at the tip of the branches. Yellow arrows indicate PTF1A⁻/GP2⁻/NKX6-1⁺ cells in the trunk of the branches. Scale bar represents 20 μm. Abbreviations: GP2, pancreatic secretory granule membrane major glycoprotein 2; NKX6-1, NK6 homeobox 1; PTF1A, pancreas specific transcription factor 1a; INS, insulin