Stress-Induced Unfolded Protein Response

Cells were stressed with various compounds for indicated times. At the start of treatment, we replaced the culture medium with DMEM/F12 medium supplemented with UPR activators tunicamycin (2.5 µg/ml; Sigma) and thapsigargin (0.33 and 1 µM; Sigma) or GCN2 activator halofuginone (10 nM; Cayman Chemical Company)^{39 (link)}, HRI activator BTdCPU (KM09748SC) (6 µM; Thermofisher)^{38 (link)}, PKR activator BEPP (10 µM; Sigma)^{37 (link)} or PERK activator CCT020312 (10 µM; Millipore) which specifically activate the ISR^{40 (link)}. All compounds were dissolved in DMSO. Control cultures were simultaneously treated with equal amounts of DMSO (vehicle control).