Characterization of Cellular and Molecular Heterogeneity of Bone Marrow Stromal Cells

Figure 4

Effect of alkaline phosphatase (ALP) gene silencing by small interfering RNA (siRNA) on a clonal population of human bone marrow stromal stem cell CL1. (A) ALP staining at day 3, day 5, or day 7 days of osteoblast differentiation (OB induction). (B) Quantitative real-time PCR for ALP gene following ALP siRNA transfection at day 0 OB and day 7 of OB. Data are presented as fold change in expression of each target gene normalized to GAPDH ( = three independent experiments, ). Western blotting analysis of day 3 and day 7 after siRNA ALP transfection of CL1 cells, ALPL specific antibody, and B-actin was used. (C) Mineralized matrix formation as visualized by Alizarin Red S staining in siRNA transfected CL1 cells after 14 days of OB induction. (D) Nile red quantification of mature lipid filled adipocyte in control noninduced (Cont), adipocyte induced (Adip I), and ALP siRNA transfected cells that are adipocyte induced (ALP siRNA). Adipocyte induction was carried out for 7 days. .