Figure 1.

Expression pattern of BMP2 in differentiating EBs. (a) Detection of the expression of BMP2 by qPCR on the samples from EBs derived from wild-type CGR8 ES cells (i) and RT-PCR (ii) on samples from EBs derived from BMP2 ES cells (for conditions, see Additional data file 14). The qPCR results are presented as the mean of three independent experiments ± standard deviation. (b) Expression of EGFP during differentiation of the BMP2 ES cells induced by the conventional hanging drop protocol. Scale bar represents 50 μm. (c) Protocol for isolation of puromycin resistant BMP2⁺ cells after treating the plated four-day-old EBs with 3 μg/ml puromycin for three days. (d,e) FACS analysis of the trypsinized untreated control and puromycin resistant BMP2⁺ cells. (f) BMP2⁺, three days after plating in gelatine-coated tissue culture dishes in the presence of 3 μg/ml puromycin. Scale bar represents 50 μm. (g,h) Detection of BMP2 in BMP2⁺ cells (g) or ES cells (h) by immunohistochemistry staining. Stainings were done after the cells were trypsinized and plated on microscopic slides for 24 hours. Scale bar represents 20 μm.