Fig. 5

Characterization of notochordal transcription factor brachyury expression via the T-fluorescent reporter construct. a Cells transfected with lentivirus containing plasmid for eGFP downstream of the brachyury (T) promoter. b Bright-field image and fluorescent image of a differentiated cell colony following culture in basal medium when supplemented with 40 ng/ml BMP4 and 20 ng/ml FGF2 (3 days) (scale bar = 100 μM). c eGFP expression detected via flow cytometry after supplementation with BMP4 and FGF2 at day 3 (showing 27.5% of cells for this population). d Before cell sorting, LV-T-GFP-positive cells were localized around the periphery of a differentiated cell colony (scale bar = 400 μM). e When cultured in basal medium supplemented with BMP4 and FGF2, flow cytometry analysis showed 25.4% of cells to be positive for T-GFP⁺ in differentiated hiPSCs. GFP green fluorescent protein, bFGF2 basic fibroblast growth factor, BMP bone morphogenetic protein 4