Ho 1 antibody

Total proteins were isolated from the lung tissue homogenates according to the protocol provided by the Protein Extraction Reagents Kit (ASPEN, South Africa). Proteins were separated on 10% SDS-PAGE gels by electrophoresis and transferred to a PVDF membrane. The membranes were blocked with 5% nonfat milk for 1 hour and probed with the following antibodies: Nrf-2 antibodies (1 : 200, Santa Cruz, California, USA), HO-1 antibodies (1 : 200, Santa Cruz, California, USA), H2A antibodies (1 : 1000, Santa Cruz, California, USA), and β-actin antibodies (1 : 2000, Merck Millipore, Germany). HP-conjugated anti-mouse IgG (Cell Signaling Technology) was used as a secondary antibody. Images were scanned with the canon imaging system and analyzed using Alpha image software.

HaCaT cells (American Type Culture Collections, Manassas, VA, USA), a human keratinocyte cell line, were cultured and maintained in Dulbecco's modified Eagle's medium (DMEM, Thermo Fisher Scientific, Inc., Waltham, MA, USA) supplemented with 1% antibiotics (penicillin/streptomycin) and fetal bovine serum (FBS, 10%) at 37°C in a 5% CO₂ humidified incubator. The HEK293-TRPV1-luciferase stable cell line (Creative Biogene Biotechnology, Shirley, NY, USA) was cultured in DMEM supplemented with 10% FBS, 10% puromycin, and 1% antibiotics at 37°C in a 5% CO₂ humidified incubator. The following are the cell culture reagents: B[a]P (CAS No. 50-32-8, purity 99.9%, Sigma-Aldrich Co., N.Y., USA), dasatinib (Src inhibitor, Sigma-Aldrich Co.), AhR antibodies (Santa Cruz Biotechnology, Santa Cruz, Ca, USA), ARNT antibodies (Santa Cruz Biotechnology), LaminB1 antibodies (Epitomic, Burlingame, CA, USA), α-tubulin antibodies (Epitomic), β-actin antibodies (Sigma-Aldrich Co.), phospho-Src (Tyr416) antibodies (Cell Signaling Technology Inc., Beverly, MA, USA), Src antibodies (Cell Signaling Technology Inc.), CYP1A1 antibodies (Santa Cruz Biotechnology), NQO1 antibodies (Santa Cruz Biotechnology), and HO-1 antibodies (Santa Cruz Biotechnology).