2.7 | Immunofluorescence Staining
Various treated LF cells were fixed in 4% PFA for 30 min at room temperature. After permeabilizing and blocking treatment, the cells were incubated with primary antibodies against a-SMA (1:500, Abcam), Gli1 (1:1000, Abcam) and WISP-1 (1:500, Abcam) overnight at 4˚C. Then, cells were washed by PBS for three times and incubated with fluorescein isothiocyanate (FITC)-conjugated goat anti-rabbit antibody (1:500, Abcam) in dark conditions. Nuclei were stained with DAPI (Beyotime, Nanjing, China) for 5 min. Immunofluorescence was observed under a fluorescence microscope (OLYMPUS, Nanjing, China).