Bs 23064r

For routine histopathological analysis, paraffin-embedded synovial tissue sections from RA and OA patients were deparaffinized and stained with hematoxylin and eosin (H&E). For immunofluorescence staining, 6-μm-thick tissue sections were incubated overnight at 4 °C with the following primary antibodies diluted in PBS: mouse monoclonal antibody against SLC7A5 (1:100, Santa Cruz, sc-374232) and rabbit polyclonal antibody to vimentin (1:100, Bioss, bs-23064R). Next morning, the samples were washed three times in PBS and incubated for 45 min at room temperature with secondary antibodies, i.e., FITC AffiniPure goat anti-mouse IgG (H+L) (1:400, Earthox, E031210-01) and Cy3 AffiniPure goat anti-rabbit IgG (H+L) (1:400, Earthox, E031620-01). 4′,6-Diamidino-2-phenylindole (DAPI) was used to detect the nucleus (1:100,000, Sigma-Aldrich, D9542). Immunofluorescent staining procedure was followed with slight modifications, as previously described [15 (link)]. The immunofluorescent images were captured with a fluorescence microscope (Olympus, Japan) and analyzed by the ImageJ software.