Fig. 3.

A Measurement of GFP2⁺ cells at d12 after subtraction of either stage 3, nicotinamide (NA), EGF, or LDN193189 (LDN). For comparison stage 4 plus protein kinase C activation by 100 nM PDBu. Data are means ± SEM, n = 3. B GFP2-expression in dependence of the EGF concentration at d12. Data are means ± SEM, n = 3. C/D Effect of different growth factors each used at 100 ng/ml on GFP2 expression (C) at d12 and pancreatic marker gene expression (D). Depicted is the relative gene expression of PDX1, SOX9, NKX6-1, and NKX2-2. Data are means ± SEM. n = 7–8 (GFP2 flow cytometry), n = 3 (RT-qPCR). (E/F) Effect of canonical Wnt-signaling on GFP2 expression (E) at d12 and pancreatic marker gene expression (F). The pathway was activated by CHIR (3 µM) or inhibited by IWP4 (1 µM) or IWR-1 (2 µM). Depicted is the relative gene expression of PDX1, SOX9, NKX6-1, and NKX2-2. Data are means ± SEM. n = 3 (GFP2, RT-qPCR). ANOVA plus Dunnett’s post-test, ***p < 0.001, **p < 0.01, *p < 0.05