Immunofluorescence Staining
CFs were cultured on sterile glass coverslips were fixed in 4% paraformaldehyde for 10 min, washed with PBS three times, and permeabilized with 0.3% Triton X-100 (Sigma-Aldrich, Saint Louis, MO) in PBS for 20 min. After blocking with 10% goat serum (Jackson ImmunoResearch, West Grove, PA) at room temperature for 30 min. Then, the cells were incubated with rabbit anti-vimentin (1:200), mouse anti-α-SMA (1:400) overnight in humidified incubator. The slides were then washed with PBS and incubated with Alexa Fluor 594 conjuncted goat anti-mouse IgG (1:400) or Alexa Fluor 488 conjuncted goat anti-rabbit (1:200; Jackson, West Grove, PA) for 30 min at 37 oC. The nuclei were counterstained with 4′,6-diamidino-2-phenylindole (DAPI, 1:1000; Sigma-Aldrich, Saint Louis, MO). Finally, the slides were mounted in fluoromount-G(Southern Biotech), and examined using a fluorescence microscope.