Aldh3a1

Small pieces of human lung samples were homogenized and lysed with cell lysis buffer and followed by measurement of protein concentrations. Equal amounts of protein were fractionated by electrophoresis and then transferred to PVDF membranes. The membranes were then incubated with antibodies against NRF2 (Santa Cruz), ALDH1A1 (Abcam), and ALDH3A1 (Santa Cruz) followed by incubation with secondary antibodies. For the detection of specific protein bands, the membranes were incubated in LumiGLO reagent and peroxide (Cell Signaling Technologies, Danvers, MA, USA), and then exposed to X-ray films.

The primary antibodies were purchased from these resources: Heme oxygenase-1 (HO-1; Santa Cruz, CA, USA, sc-136960), superoxidase dismutase 2 (SOD2; Santa Cruz, CA, USA, sc-133134), aldehyde dehydrogenase (ALDH) 1A1 (Santa Cruz, CA, USA, sc-374076), ALDH2A1 (Santa Cruz, CA, USA, sc-100496), ALDH3A1 (Santa Cruz, CA, USA, sc-376089), and GAPDH (Santa Cruz, CA, USA, sc-47724). The secondary antibodies were horseradish peroxidase-conjugated anti-mouse IgG (Cell Signaling Technology, 7076) and anti-rabbit IgG (Cell Signaling Technology, 7074). An Annexin V-FITC Apoptosis Detection kit was purchased from eBioscience (San Diego, CA, USA). Dimethylsulfoxide (DMSO, # 41639), 1,1-Diphenyl-2-picryl-hydrazyl (DPPH), and ascorbic acid were purchased from Sigma-Aldrich (St. Louis, MO, USA). GSH-Glo assay kit (Promega, Madison, WI, USA, # V6911) and ROS-Glo™ H₂O₂ assay kit (Promega, Madison, WI, USA, #G8820) were purchased from Promega (Madison, WI, USA, V6911).