Fig. 6. Osteogenic differentiation study on rASCs (A) representative western blot result. ALP, OCN expressions were evaluated during 4 weeks. (B) The relative expressions of osteogenic marker protein were calculated. In weeks 2 and 3, ALP expression was increasingly promoted with increased RGO, indicating that the more RGO is incorporated, the faster the early stage of osteogenesis is. In weeks 3 and 4, OCN expression of 1% RGO was highly increased, indicating that 1% RGO underwent late stage osteogenesis earlier. (C) ICC result. OCN expression for 4 weeks is depicted. At 4 weeks, 1% RGO had the highest expression. Scale bars = 100 μm. (D) Three dimensional images of OCN expression on 1% RGO. Hemisphere-shaped cell aggregation indicates higher OCN expression. (E) Representative images of ARS. (F) Quantitative analysis of ARS. The calcium deposition rate of 1% RGO in week 4 had the highest value with a significant difference. (n = 5, LSD) ALP: alkaline phosphatase; OCN: osteocalcin, ICC: immunocytochemistry. ARS: alizarin red staining.