Anti goat dylight 594 secondary antibody

Chondrogenic differentiation was performed according to^{105 (link)} with minor modifications. Briefly, 5 × 10⁴ WT and mutant MenSCs were left aggregated in microwell plates and then provided with chondrogenic medium containing high-glucose DMEM, 10% FBS, 10 μg/ L TGF-β3, 0.1 μmol/ L dexamethasone, 50 μmol/ L vitamin C, and 6.25 mg/L insulin. The medium was changed every 3–4 days. Control cells were kept in a regular culture medium. After 20 days of induction, cells were fixed in 4% FA and incubated with the goat anti-human aggrecan antigen affinity-purified polyclonal antibody (1:500; R&D, cat# 967,800) followed by incubation with anti-goat DyLight™ 594 secondary antibody (1:500) and 1 μM Hoechst 33,342 (Life Technologies).

Adipogenic differentiation was performed according to^{104 (link)} with minor modifications. Briefly, WT and mutant MenSCs at passages 4–7 were plated at a density 20,000 cells/cm² in a 12-well plate in a regular culture medium. At 90–100% confluence, the culture medium was replaced by adipogenic induction medium, including high-glucose DMEM, 10% FBS, 0.5 mM 3-isobutyl-1-methylxanthine (Sigma, cat # I5879), 100 µM indomethacin (Sigma, cat # I7378), 0.1 µM dexamethasone and 10 µg/ mL insulin. Control cells were kept in regular culture medium. After 20 days of induction, cells were fixed in 4% FA and incubated with the goat anti-mouse fatty acid-binding protein 4 (FABP4) antigen affinity-purified polyclonal antibody (1:500; R&D, cat# AF3150), followed by incubation with anti-goat DyLight™ 594 secondary antibody (1:500) and 1 μM Hoechst 33,342 (Life Technologies).