Relative alkaline phosphatase activity (a) and mRNA expression of TNAP ((b) qRT-PCR) in various stem/progenitor cells. Bone marrow-derived MSC (bmMSC) has both high AP activity and TNAP expression, followed by ES cells. Relative expression (qRT-PCR) of common pluripotent (Oct-4 (c) and Nanog (d)) and osteogenic (Osteocalcin, OstC (e) and Osteopontin, OstP (f)) genes. A high level of Oct-4 and Nanog documented the real pluripotent status of ES cells. A higher expression of OstC and OstP mRNA in bmMSC documented their osteogenic properties/lineages specification. (ES: mouse embryonic stem cells; NSC: neural stem/progenitor cells; bmMSC: bone marrow mesenchymal stem cells; aMSC: adipose tissue mesenchymal stem cells). Details of the presented assay may be found in our previous work [9, 77]. Primers and conditions for OstC and OstP were as follows: OstC: 5′-CTTGGGTTCTGACTGGGTGT-3′, 5′-GCCCTCTGCAGGTCATAGAG-3′ (212 bp, 60°C); OstP: 5′-TCACCATTCGGATGAGTCTG-3′, 5′-ACTTGTGGCTCTGATGTTCC-3′ (436 bp, 60°C). Error bars indicate ±SD (n ≥ 3, *
P < 0.05; **
P < 0.01, ANOVA post hoc Bonferroni's Multiple Comparison Test).