Fig. 4

In vitro osteogenic differentiation in HUVECs after OCT4-30Kc19 and BMP4 treatment. (A) Quantitative polymerase chain reaction (qPCR) for osteogenic marker genes, COL I, OPN, and BSP. HUVECs were treated with OCT4-30Kc19 and BMP4 for 2 weeks in osteogenesis medium. PCR analysis showed elevation in osteogenic markers when HUVECs were treated with OCT4-30Kc19 and BMP4 (n = 3). ^*p < 0.05, ^**p < 0.01, ^***p < 0.001. (B) Alizarin red s staining (ARS) showing calcium deposition in HUVECs. The sample treated with OCT4-30Kc19 and BMP4 showed the highest calcium deposition. Quantitative analysis of ARS (n = 3). ^***p < 0.001. (C) Immunofluorescence staining of OCN in HUVECs. OCN was expressed in HUVECs treated with OCT4-30Kc19 and BMP4. Anti-OCN-antibody (1:400) was used as the primary antibody for OCN detection. Blue and green represent the nucleus and OCN, respectively. Scale bar, 50 μm. Mean fluorescence intensity (MFI) of each group was quantified with Image J software (n = 3). ^*p < 0.05, ^**p < 0.01