Gene-expression analysis of SSEA-1⁺ cells by QRT-PCR. (A) Representative gel electrophoresis of RT-PCR products from different genes using GAPDH as an internal PCR control. Different cell types were used as positive controls to assure the specificity of the PCR reaction. Differentiated ES cells for Brk (T; Brachyury) and Gos (Goosecoid); SSEA-1⁺–derived myoblasts for PDGF-Rα and Tbx6; SSEA-1⁺–derived endothelial-like cells for KDR; CD45⁺/KLS HSCs for VE-cadherin, Runx1, Scl/Tal1, Gata-2 (exons 5/6), Gata-2 (1S exon/1S intron), Gata-2 (1S exon/exon 2), c-Kit, and Slam1; and SSEA-1⁺–derived hepatocyte-like cells for E-cadherin and Sox17 were used, respectively. (B) The values indicate the relative mRNA expressions using the ΔCT value of Brachyury as a reference. All quantitative results shown are mean values from 2 independent experiments, each performed in duplicate. The different mesenchymal populations used in this study were clone-1 (Cl1), clone-2 (Cl2), clone-3 (Cl3) SSEA-1⁺ cells, bulk SSEA-1⁻ cells (S⁻), and both SSEA-1⁺ (S⁺) and SSEA-1⁻ fractions isolated from CD45/CD11b/Ter119/CD31⁻ bone marrow fraction.