Tcl sl

hMSCs were fixed with 3.7% paraformaldehyde in PBS, followed by permeabilization with -20°C acetone, and incubated with 1% bovine serum albumin to block nonspecific binding of antibodies. For N-cadherin and fibronectin staining, the specimens were incubated with respective primary antibodies (Abcam, UK) diluted at 1 : 300 for 1 h and with appropriate FITC secondary antibodies (Abcam, UK) diluted at 1 : 600 for 1 h. Nuclei were stained by Hoechst (Molecular Probes, USA) in blue. The fluorescently stained samples were imaged by using a laser scanning confocal microscopy system (Leica TCL SL, Germany).

The cells were fixed with 3.7% paraformaldehyde followed by a permeabilization process using cold acetone. The cells were then blocked with 1% bovine serum albumin for 30 min, and incubated with N-cadherin or fibronectin antibody (Abcam, Cambridge, UK) at 1:300 dilutions in phosphate buffered saline (PBS) for 1 h at room temperature. The cells were then washed in PBS, incubated with fluorescein isothiocyanate (FITC) secondary antibodies (Abcam, Cambridge, UK) at a concentration of 1:600 dilution for 1 h in the dark, and counterstained for nuclei with Hoechst (Molecular Probes, Oregon, USA) for 10 min. Stained cells were analysed under a laser scanning confocal microscope (Leica TCL SL, Germany).