Fig. 4

CG-stimulated ASCs showed chondrogenic characteristics. a Expression of chondrogenic differentiation markers in ASC micromass cultures. Total RNA was extracted from ASC micromass cultures at 14 days. Expression of each marker mRNA was examined using RT-PCR. b Chondrogenic ECM staining of ASC micromass cultures. Sections of ASC micromass cultures at 21 days were stained with Alcian Blue or Safranin O. c Expression of ACAN and COL2A1 proteins in ASC micromass cultures. Sections of ASC micromass cultures at 21 days were stained with a fluorescence (phycoerythrin)-conjugated anti-ACAN or COL2A1 antibody (red: ACAN or COL2A1, blue: DAPI). ASCs were stimulated with CG (2400 g for 30 min) or TGF-β1 (10 ng/mL) and then used for micromass culture. The control was not stimulated with CG or TGF-β1. ACAN aggrecan, CG centrifugal gravity, COL1 collagen type I, COL10 collagen type X, COL2A1 collagen type II ​alpha 1, CON control, TGF transforming growth factor