Dimethylmethylene blue dye

Reagents were obtained from Sigma (Munich, Germany) unless otherwise indicated. Recombinant FGF-2 and TGFβ3 were purchased from Peprotech (Hamburg, Germany). The dimethylmethylene blue dye was from Serva (Heidelberg, Germany). The anti-IGF-I (AF-291-NA) antibody was from R&D Systems (Wiesbaden-Nordenstadt, Germany), the anti-type-II collagen (II-II6B3) antibody from the NIH Hybridoma Bank (University of Iowa, Ames, IO, USA), the anti-type-I (AF-5610) antibody from Acris (Hiddenhausen, Germany), the anti-bromodeoxyuridine (BrdU; BU-33) and anti-type X collagen (COL-10) antibodies from Sigma, and the anti-SOX9 (C-20), anti-CD34 (C-18), anti-CD71 (C-20), and anti-CD105 (T-20) antibodies from Santa Cruz Biotechnology (Heidelberg, Germany). Biotinylated secondary antibodies and ABC reagent were obtained from Vector Laboratories (Alexis Deutschland GmbH, Grünberg, Germany). The IGF-I enzyme-linked immunosorbent assay (ELISA) (hIGF-I Quantikine ELISA) was from R&D Systems, the type II and type I collagen ELISAs (Arthrogen-CIA Capture ELISA Kit) from Chondrex (Redmond, WA, USA), and the type X collagen ELISA from Antibodies-online GmbH (Aachen, Germany). The Cell Proliferation reagent WST-1 was from Roche Applied Science (Mannheim, Germany) and the alkaline phosphatase (ALP) staining kit was from Sigma.

All reagents were purchased at Sigma (Munich, Germany) unless otherwise indicated. The dimethylmethylene blue dye was from Serva (Heidelberg, Germany). Recombinant TGF-β3 was from Peprotech (Hamburg, Germany). The antibodies used for immunohistochemical analyses were as follows: the anti-β-galactosidase (β-gal) (GAL-13) and anti-type-X collagen (COL-10) antibodies from Sigma, the anti-TGF-β (V), anti-SOX9 (C-20), and anti-FLAG tag (BioM2) antibodies from Santa Cruz Biotechnology (Heidelberg, Germany), the anti-type-I collagen (COL-1) antibody from Abcam (Cambridge, UK), and the anti-type-II collagen (II-II6B3, NIH Hybridoma Bank, University of Iowa, Ames, USA) antibody from Acris (Hiddenhausen, Germany). Biotinylated secondary antibodies and the ABC reagent were purchased at Vector Laboratories (Alexis Deutschland GmbH, Grünberg, Germany). The TGF-β enzyme-linked immunosorbent assay (active hTGF-β1 Quantikine ELISA) was from R&D Systems (Wiesbaden, Germany).

All reagents were from Sigma (Munich, Germany) unless otherwise indicated. Recombinant FGF-2 and TGF-β3 were purchased at Peprotech (Hamburg, Germany). The dimethylmethylene blue dye was from Serva (Heidelberg, Germany). The anti-TGF-β (V) and anti-SOX9 (C-20) antibodies were from Santa Cruz Biotechnology (Heidelberg, Germany), the anti-type II collagen (II-II6B3) antibody from the NIH Hybridoma Bank (University of Iowa, Ames, IA, USA), the anti-type I collagen (AF-5610) antibody from Acris Antibodies (Hiddenhausen, Germany), and the anti-type X collagen (COL-10) antibody from Sigma. Biotinylated secondary antibodies and the ABC reagent were purchased at Vector Laboratories (Alexis Deutschland GmbH, Grünberg, Germany). The TGF-β enzyme-linked immunosorbent assay (hTGF-β1 Quantikine ELISA) was from R&D Systems (Wiesbaden, Germany).