Igg rabbit polyclonal antibody

NHEKs were cultured on slides (Lab-Tek, Rochester, NY, USA). These slides were then washed in phosphate-buffered saline (PBS), fixed with acetone for 10 min, and blocked using 10% bovine serum albumin (Roche Diagnostics, Basel, Switzerland) in PBS for 30 min for the staining of IL-37. Samples were incubated with primary anti-human IL-37 polyclonal rabbit antibody (1:100) (Abcam) or IgG rabbit polyclonal antibody (Abcam) in Western Breeze Blocker/Diluent (Thermo Fisher Scientific) overnight at 4°C. The slides were then washed with PBS before incubation with anti-rabbit secondary antibody (Alexa Fluor 546; Molecular Probes, Eugene, OR, USA) for 1 h at room temperature. After nuclear staining with 4′,6-diamidino-2-phenylindole (DAPI), the slides were mounted with UltraCruz mounting medium (Santa Cruz Biotechnology, Dallas, TX, USA). All samples were analyzed using a D-Eclipse confocal laser scanning microscope (Nikon, Tokyo, Japan).

Tapinarof (MedChemExpress, Monmouth Junction, NJ, USA) was dissolved in dimethyl sulfoxide (DMSO; Nacalai Tesque, Kyoto, Japan) and stored at −80°C until used in the experiments. GFF was obtained from P&G Innovation Godo Kaisha (Kobe, Japan). Anti-human IL-37 polyclonal goat antibody (R&D Systems, Minneapolis, MN, USA), anti-human IL-33 monoclonal mouse antibody, anti-human IL-36γ/IL-1F9 monoclonal mouse antibody (Abcam, Cambridge, UK), anti-phosphorylated ERK-1/2 rabbit monoclonal antibody (Thr202/Tyr204), anti-ERK-1/2 rabbit monoclonal antibody, anti-phosphorylated p38 rabbit monoclonal antibody (Thr180/Tyr182), anti-p38 rabbit monoclonal antibody, anti-phosphorylated JNK rabbit monoclonal antibody (Thr183/Tyr185), anti-JNK rabbit polyclonal antibody, anti-AHR monoclonal rabbit antibody, and anti-human β-actin monoclonal mouse antibody (Cell Signaling Technology, Danvers, MA, USA) were used for western blotting. Anti-human IL-37 polyclonal rabbit antibody and IgG rabbit polyclonal antibody (Abcam) were used for immunofluorescence.