Tris glycine gel

For protein extraction, the cells were sonicated on ice in RIPA Buffer (Biosesang, Seongnam, Korea) containing a protease inhibitor cocktail. After centrifugation at 12,000 rpm at 4°C for 20 min, clear cell lysates were measured for protein concentration by Bradford assay. A total of 20 μg protein was fractionated by SDS-PAGE on 8–16% Tris- glycine gel (Komabiotech, Seoul, Korea), transferred to nitrocellulose membrane (Invitrogen), and then blotted with antibodies against LC3 II (1:500), p62/SQSTM1 (1:4000), beclin 1 (1:10000), ATG5 (1:500), Nrf2 (1:1000) (Novus biological, Littleton, USA), pro-IL-1β (1:1000), p38 (1:1000), phospho-p38 (pp38, 1:1000) (Cell Signaling Technology, Danvers, MA), or β–actin (1:200, Santa Cruz Biotechnology).

For protein extraction, tissues were lysed with a sonicator in RIPA Buffer (Biosesang, Seongnam, Korea) including a protease inhibitor cocktail (Thermo Fisher Scientific, Waltham, MA). Protein concentration was measured by Bradford assay. 50 µg protein was fractionated by SDS-PAGE on 8-16% Tris-glycine gel (Komabiotech, Seoul, Korea), transferred to methanol-presoaked PVDF membrane (Invitrogen, Waltham, MA), and blotted with antibodies against H2 antigen (1:500, Cat# sc-59467, Santa Cruz Biotechnology), FUT1 (1:500, Cat# sc-21963, Santa Cruz Biotechnology), and β–actin (1:1000, Cat# sc-47778, Santa Cruz Biotechnology).