Western Blot Analysis of A549 Cells

WB was performed according to the previously reported method.⁴⁷ (link) Proteins were extracted from A549 cells using a sample buffer (Nacalai Tesque), and the lysates were denatured at 95°C for 3 min. The lysates were then resolved by SDS-PAGE (SuperSep Ace gel; FUJIFILM Wako, Osaka, Japan) and transferred to a polyvinylidene difluoride membrane (Pall Corporation, Port Washington, NY, USA). Antibody reactions were performed using the Can Get Signal immunoreaction enhancer solution (Toyobo, Osaka, Japan). Immunoreactivity was visualized using Chemi-Lumi One Super (Nacalai Tesque) or ImmunoStar LD (FUJIFILM Wako) and ChemiDoc MP imaging system (Bio-Rad, Hercules, CA, USA). The antibodies used in this study are listed in the Antibody List.