Figure 3: Immunofluorescence analysis of the C-peptide expressing cells. (a) Insulin-producing cells stain positive for C-peptide expression (C-peptide stain, green; DAPI stain, blue). RAW 264.7 (Mouse leukaemic monocyte macrophage cell line) is the negative control. Scale bars = 200 μm. (b) Experimental time course for the differentiation of ES-D3 cells to IPCs. For the quantification of IPCs, at least 10 images for each treatment were taken using an EVOS fluorescent microscope (USA). Total cell number was quantified based on DAPI nuclear staining and C-peptide expressing cells were quantified using Image J software (NIH, US). Bars represent means ± SD from three independent experiments. * significantly different from 7 days.