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. 2021 Jun 7;12:3330. doi: 10.1038/s41467-021-23525-x

Fig. 3. Systematic screening for the differentiation of pancreatic progenitors (PPs) into β-cells.

Fig. 3

a A scheme of inducing PP-10C-treated PPs into β-cells using the improved protocol. There are extra five differentiation stages from hPSCs to PP-10C-treated PPs, so these three late stages are named as stage-6, -7, and -8, respectively. Chemicals/factors highlighted in red have not been reported in literatures for inducing hPSCs into β-cells. The chemicals/factors used in stage-6, -7, and -8 are termed as EP-8C, iβ-9C, and Fβ-7C, respectively. b Cell cytometry analysis of stage-8 cells showed Fβ-7C collectively induced (NKX6.1+/INS+) β-cells from earlier stages, while subtracting some factors from Fβ-7C at stage-8 reduced the efficiency (n = 5). c, d 10C-treated PPs were induced to β-cells according to the Method 3 shown in Fig. 3a, followed by immunostaining in the end-point cells, showing robust expression of INS, NKX6.1 but not GCG. Figure 3c was stitched by individual images. The inserted small image on the right bottom of Fig. 3c was the DAPI staining. Scale bar, 50 µm for the right panel of 2D and 100 µm for all others. Images of c and d are representative of five independent experiments, respectively.