Figure 2

Mesodermal differentiation of mSSCs. (A, B) mRNA profiles of pluripotency-related marker genes (A: Oct3/4, Nanog) and mesoderm lineage - specific genes (B: Brachyury, Flk1, Mesp1, Islet1, Nkx2.5) in mSSCs and ESCs during in vitro vascular differentiation process. Gene expression was measured using real-time RT-PCR and normalized as its relative ratio to GAPDH. The mRNA levels are expressed relative to the corresponding mRNA level at day 0 (undifferentiated mSSCs and ESCs; set as 1). The data were obtained from 3 independent preparations and are expressed as mean ± SEM. (C, D) Representative FACS histograms and immunofluorescent images showing Flk1 expression in differentiated mSSCs (diff. mSSCs) and differentiated ESCs (diff. ESCs). Approximately 25% and 30% of differentiated cells from mSSCs and ESCs on day 9 of vascular differentiation were positively stained with anti-Flk1 IgG. Murine ECs were included as a positive control. Each histogram in (C) is representative of three separate experiments. In addition, cells on day 14 were stained with anti-Flk1 or nonspecific control IgGs. Scale bar is 50 µm. Nuclei were stained with DAPI.