Nanog pe

FCM analysis of the P4 BM-MSC preparations was performed with a FACScan instrument (BD Biosciences, Franklin Lakes, NJ, USA) using BD CellQuest Pro software. Cells were stained at 4 °C for 30 min with the following primary antibodies: SSEA-4, TRA-1-60, and TRA-1-81 for detecting ESC surface markers (all of the above antibodies were purchased from Millipore). For the intracellular staining, the cells were fixed, permeabilized, and stained with SOX2-PE (Becton Dickinson), NANOG-PE (R&D Systems), and anti-human OCT3/4 primary antibody, respectively. The cells were washed and then stained with FITC-conjugated goat anti-mouse secondary antibody (GAM-FITC, purchased from Becton Dickinson) and FITC-conjugated goat anti-rat IgG (KPL, USA) at 4 °C for 30 min with light protection. Mouse IgG1-PE and Mouse IgG1-FITC (both were purchased from Becton Dickinson, USA) were served as an isotype control.