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).