Transcription factor SPB-x is a key molecule inducing hypertrophy of differentiated chondrocyte from MSC G.-I. Im, J.-M. Lee, J.-M. Ahn, E.-A. Kim Osteoarthritis.

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Transcription factor SPB-x is a key molecule inducing hypertrophy of differentiated chondrocyte from MSC G.-I. Im, J.-M. Lee, J.-M. Ahn, E.-A. Kim Osteoarthritis and Cartilage Volume 22, Pages S27-S28 (April 2014) DOI: 10.1016/j.joca.2014.02.073 Copyright © 2014 Terms and Conditions

Figure 1 Chondrogenic potential of lenti-miR-im6 transduced BMSCs. (A) Expression pattern of miR-im6 in presence of PTHrP for chondrogenic differentiation. (B) Transduction efficiency of the produced miR-im6 lentivirus in BMSCs. (C) Ratio of GAG/DNA measured in cultured lentiviral transduced pellets after 4 weeks of culture. The bars represent means ± SD. CM: BMSCs treated with their respective chondrogenic medium. CMT: BMSCs treated with chondrogenic medium containing 5 ng/ml TGF-β. CMTP: BMSCs treated with chondrogenic medium containing 5 ng/ml TGF-β and 100 ng/ml PTHrP (D and F) Gross appearances and Safranin-O staining of each chondrogenic pellet. (E) Protein expression of chondrogenic differentiation and hypertrophic markers in the lentiviral transduced pellets. Detection of type II collagen, SOX9, ALP, and type X collagen at 4 week after chondrogenic differentiation using Western blotting. Osteoarthritis and Cartilage 2014 22, S27-S28DOI: (10.1016/j.joca.2014.02.073) Copyright © 2014 Terms and Conditions

Figure 2 Expression pattern of major signals by SPB-X down- or up-regulation in hBMSCs. (A and B) Genes and proteins expression in lenti-im6 transduced hBMSCs. (C) Construct of SPB-X overexpression vector. (D) Fluorescent microscopy of hSPB-X overexpressed BMSCs. (E and F) Rq-PCR and Western blotting results of hSPB-X overexpressed BMSCs. Osteoarthritis and Cartilage 2014 22, S27-S28DOI: (10.1016/j.joca.2014.02.073) Copyright © 2014 Terms and Conditions

Figure 4 Effect of SPB-X knockout in chondrogenic differentiation of mouse MSCs. (A) Ratio of GAG/DNA measured in mouse MSC pellets after 3 weeks of culture. (B) Gross appearance of mouse MSCs chondrogenic pellets. (C) Protein expression of major hedgehog signal molecules, chondrogenic differentiation and hypertrophic markers in mouse MSCs chondrogenic pellets. (D) Blocking mechanism of chondrocyte hypertrophy by miR-im6. Osteoarthritis and Cartilage 2014 22, S27-S28DOI: (10.1016/j.joca.2014.02.073) Copyright © 2014 Terms and Conditions