Figure 4. Skewed differentiation was induced in IHH-depleted BMSC. (Aa, b), (C and D) BMSC (n = 5) were transfected with siRNA negative control or siRNA IHH for 24hours then incubated in adipogenesis differentiation medium for 21 days. Adipocytes were visualized under inverted light microscope and adipogenesis measured by staining fat droplets using Oil-Red-O stain. (Ba, b), (E, F) BMSC (n = 5) were transfected with siRNA negative control or siRNA IHH for 24hours then incubated in osteogenic differentiation medium for 21 days. Osteogenesis and Ca2 mineralization measured using Alizarine-Red-S stain. (G) RT-PCR for adipogenesis markers, PPARγ, LPL, and FABP4, and osteogenesis markers, ALP-BMA, RUNX2, and osteocalcin in BMSC with and without IHH siRNA. GAPDH was used as a housekeeping gene. Results have shown as mean ± SEM. *P <0.05, **P < 0.01, ***p <0.001.