Modulation of GRP78 Signaling in Primary Murine Macrophages

Primary MC from C57Bl/6 mice were isolated for culture using Dynabeads. They were cultured in DMEM (1,000 mg/L or 5.6 mM glucose) supplemented with 20% FBS, 100 µg/mL streptomycin, and 100 µg/mL penicillin at 37°C in 95% O₂, 5% CO₂. Cells were serum-deprived in 0.5% FBS for 24 h prior to treatment with HG (30 mM) with or without inhibitors of csGRP78 signaling: the C-terminus targeting GRP78 antibody C38 (2 µg/mL) (Munro and Pelham, 1987 (link); De Ridder et al., 2012 (link); Trink et al., 2022 (link)) or vaspin (100 ng/mL) (Nakatsuka et al., 2012 (link); Nakatsuka et al., 2013 (link); Abdolahi et al., 2022 (link)), or the following PI3K/Akt inhibitors: wortmannin (100 ng/mL), LY294002 (20 µM), and Akt Inhibitor VIII (10 µM).