Bdp 13176

Rat IL-1β protein (purity > 90%) was acquired from Sino Biological company (Beijing, China). RNA interference sequences for ADAM8 were purchased from GenePharma in Suzhou, China. Thermo Fisher Scientific (St. Louis, USA) provided DMEM/F12 for cell culture, as well as fetal bovine serum (FBS) and phosphate buffer saline (PBS). Monosodium iodoacetate (MIA) was obtained from Sigma in Gillingham, UK. Antibodies included anti-MMP-13, TNF-α, IL-6, COX2, P-JNK, JNK, P-ERK, ERK, P-P38, P38, and β-actin antibodies were obtained from Abcam, UK. The antibodies included anti-ADAM8 and MMP-1 were obtained from ThermoFisher, USA. BDP-13176, a FSCN1 inhibitor, was purchased from MedChemExpress (New Jersey, USA).

Fscn1 inhibitors NP-G2-044 (Selleckchem, Houston, TX, USA) and BDP-13176 (MedChemExpress, Monmouth Junction, NJ, USA) were reconstituted in DMSO (Roth, Karlsruhe, Germany). APC-eFl70-labeled anti-CD11c (clone N418), FITC-MHCI (28148), eFl450-MHCII (M5/114.15.2), APC-CD40 (1C10), PerCP-eFl710-CD80 (16-10A1), PE/Cy7-CD86 (GL-1), PE/TexasRed-CD274/PD-L1 (10F.9G2), PE-CD273/PD-L2 (Ty25), eFl506-CD3 (145-2C11), SB600-CD11b (M1/70), SB702-CD19 (eBio103), PE-NK1.1 (PK136), PE-eFl610-Ly6G (1A8-L6g), eFl780-FVD and eFl450-FVD used for flow cytometric analysis were purchased from BD Biosciences (Franklin Lakes, NJ, USA), BioLegend (San Diego, CA, USA) or ThermoFisher (Waltham, MA, USA). Unlabeled mouse anti-human Fscn1 antibody (clone 55K2; Sigma-Aldrich, Deisenhofen, Germany), a corresponding isotype control antibody (mouse IgG1, clone MOPC-21, BioLegend), secondary AF488-labeled IgG goat anti-mouse antibody (948492), AF647-labeled anti-CD11c antibody (clone N418), Hoechst (nuclear staining) and Alexa Fluor 555 phalloidin (F-actin) (all from ThermoFisher) were used for confocal laser scanning analysis (CLSM).