Figure 4

Immunofluorescence staining of Brachyury and Oct3/4 in ES-D3 cells treated with GSK3 inhibitors. (A) Fluorescence micrographs showing the protein expression of Brachyury/Oct3/4 after a three day differentiation of ES-D3 cells with BIO (0.5 μM), SB-216763 (5 μM), CHIR-99021 (5 μM), or CHIR-98014 (1 μM). Nuclei were counterstained with DAPI (blue). The merge is shown in yellow. Original magnification 400x. Scale bar = 50 μm. Quantitative analysis of Brachyury-positive cells after a three day treatment with of 0 – 10 μM CHIR-99021 (B) and 0 – 2.5 μM CHIR-98014 (C). Values are means ± SEM of six analyzed images of 2 different experiments.