Figure 3.

ciGEnCs synthesize CS and HA in vitro. ciGEnCs, grown on 96-well plates were treated with either vehicle or 1 mg/mL hyaluronidase for 1 hour at 37°C (A–C) or with 1 mU/mL chondroitinase or 1 mU/mL heat-inactivated chondroitinase for 2 hours at 37°C (D–F). Cells were then washed extensively and a portion was fixed (A and D), with the remainder left for a further 24 hours in serum-free medium (B, C, E, and F). These cells were then fixed, stained for HA (A–C) or CS (D–F), and counterstained with DAPI. The fluorescence intensity of HA (G) and CS (H) was quantified using a plate reader and was normalized to that of DAPI. Data were analyzed using one way ANOVAs (overall P < 0.0001) and expressed as means ± SEM. n = 8. Bonfferoni post hoc tests (versus vehicle) indicated on graphs. ^∗P < 0.05, ^∗∗P < 0.01, and ^∗∗∗P < 0.001.