Fig. 4

Characterization of endocrine markers in hPSC-derived MPCs. Immunofluorescence images of hPSC-derived MPCs using protocol 2 showing the expression of chromogranin A (CHGA; green) and NKX6.1 (red) in monolayer culture (a, b) and 3D aggregates (c). High magnification images showed that CHGA and NKX6.1 (arrowheads) were not colocalized in the same cells (b). The 3D aggregates showed no expression of CHGA, an early endocrine marker (c). d Immunofluorescence images of hPSC-derived MPCs showing expression of CHGA (green) and NKX2.2 (red). NKX2.2 was noticed in the 3D aggregates in the absence of CHGA, but both were expressed in the area surrounding the 3D aggregates. e Immunofluorescence images of hPSC-derived MPCs showing expression of neurogenin 3 (NGN3; green) and NKX6.1 (red). Note the absence of NGN3 in NKX6.1⁺ cells (arrowheads). f Immunofluorescence images of hPSC-derived MPCs showing expression of CHGA (red) and PDX1 (green). Note the presence of CHGA in the same areas expressing PDX1. All data shown are representative results from at least three independent experiments. Scale bars (a, c) = 100 μm and (b, d, e, f) = 50 μm