Zf 0312

HGFs were immersed in acetone and washed with PBS. Then, HGFs were fixed in 4% paraformaldehyde at 4°C overnight and washed with PBS three times. The following primary antibodies were used for cell incubation at 37°C for 2 h: Vimentin (1:100; ZM-0260; Beijing Zhongshan Golden Bridge Biotechnology Co., Ltd., Beijing, China) and S100A₄ (1:100; ZA-0257; Beijing Zhongshan Golden Bridge Biotechnology Co., Ltd.). H₂O₂ (3%) was added to blocking protease for 20 min at room temperature and then 30 µl sheep serum blocking buffer (ZC-02125; Beijing Zhongshan Golden Bridge Biotechnology Co., Ltd.) was added for 1 h at room temperature. Following three washes with PBS, the cells were incubated with the following secondary antibodies at 37°C for 1 h: Fluorescein isothiocyanate-conjugated goat anti-mouse immunoglobulin G (IgG; 1:100, ZF-0312; Beijing Zhongshan Golden Bridge Biotechnology Co., Ltd.) and rhodamine-conjugated goat anti-rabbit IgG (1:100; ZF-0316; Beijing Zhongshan Golden Bridge Biotechnology Co., Ltd.). Cell nuclei were stained with 4′,6-diamidino-2-phenylindole for 10 min at room temperature. Finally, the cells were photographed under a laser confocal microscope (Olympus Corporation, Tokyo, Japan).