Clone cat 5h10

After stimulation, cells were fixed with 4% paraformaldehyde for 5 min at room temperature, permeabilized with 0.01% Triton X-100 for 5 min, and blocked with 5% BSA in PBS for 20 min. Specimens were incubated with primary mouse anti-IDO1 (1:500, clone CAT-5H10, Thermo Fisher Scientific, Waltham, MA, USA) and an anti-αSMA antibody (1:200) at room temperature, followed by the secondary AlexaFluor555 goat anti-rabbit antibody (1:8000) at room temperature for 1 h. Hoechst 3342 (1:1000) was used to label the nuclei. Immunofluorescence was analyzed using a ZEISS Axio Scope5/ A1 fluorescence microscope.

After stimulation, cells were fixed with 4% paraformaldehyde for 5 min at room temperature, permeabilized with 0.01% Triton X-100 for 5 min and blocked with 5% BSA in PBS for 20 min. Specimens were incubated with primary mouse anti-β-catenin (1:500, clone CAT-5H10, Thermo Fisher Scientific, Waltham, MA, USA) and an anti-αSMA antibody (1:200, clone 1A4, Biolegend, San Diego, CA, USA) at room temperature, followed by the secondary AlexaFluor555 goat anti-mouse antibody (1:8000) and phalloidin (1:1000, all Thermo Fisher Scientific) at room temperature for 1 h. Hoechst (1:1000, Thermo Fisher Scientific) was used to label the nuclei. Immunofluorescence was analyzed using Leica DM5500 B fluorescence microscope with DFC365 FX camera (Leica Microsystems, Wetzlar, Germany).