Fig. 4

MDLS reinforced the pluripotency of P19 cells. A The alkaline phosphatase activity of MDLS-induced colonies in adherent culture. B The alkaline phosphatase activity of MDLS-induced EBs in suspension culture. C The expression of Oct4, Sox2, and Nanog (the pluripotency markers) in MDLS-induced colonies in adherent culture. Left: P19 cells were treated with MDLS (5 μg/mL) or DMSO for 48 h. Subsequently, the expression of Oct4, Sox2, and Nanog in the colonies was detected using Western blot analysis. Right: quantitative analysis of Western blotting data by image J soft-ware. Levels of these proteins were normalized to the corresponding GAPDH band expressed as relative fold changes in comparison to control samples. Values correspond to the mean ± S.E. of three independent experiments. D The expression of Oct4, Sox2, and Nanog (the pluripotency markers) in MDLS-induced EBs detected by western blot analysis. Left: P19 cells were plated onto 0.5% agarose-coated six-well plates and treated with MDLS (5 μg/mL) or DMSO for 5 days. The expression levels of Oct4, Sox2, and Nanog were detected by western blot analysis in embryoid bodies. Right: quantitative analysis of Western blotting data by image J soft-ware. Levels of these proteins were normalized to the corresponding GAPDH band expressed as relative fold changes in comparison to control samples. Values correspond to the mean ± S.E. of three independent experiments. E The expression of Oct4 and Nanog (the pluripotency markers) in MDLS-induced EBs detected by immunofluorescence assay