Hydroxychloroquine hcq

Bafilomycin A₁ (Baf), puromycin, blasticidin S and Hanks' balanced salt solution (HBSS) were purchased from FUJIFILM Wako Pure Chemical Corporation. Rapamycin was obtained from LC Laboratories. Hydroxychloroquine (HCQ) was obtained from Cayman Chemical Company. The GAK inhibitor (GAKi; cat. no. 538770), Y-27632 (cat. no. 688000) and cycloheximide were purchased from Calbiochem (Merck KGaA). DAPI was obtained from Sigma-Aldrich (Merck KGaA). Protease inhibitor cocktail and phosphatase inhibitor cocktail were obtained from Nacalai Tesque, Inc. Baf and Rapamycin were dissolved in DMSO and used at a concentration of 10 nM, and the same amount of DMSO was used for negative control. HCQ was dissolved in water and used at a concentration of 50 µM. GAKi was dissolved in DMSO and used at the indicated concentrations (3, 10 or 30 µM), and the same amount of DMSO was used for negative control. Y-27632 was dissolved in water and used at a concentration of 10 µM. Cycloheximide was dissolved in DMSO and used at a concentration of 30 µM. Cells treated with Baf (8 or 24 h), Rapamycin (24 h), HCQ (8 or 24 h), GAKi (8 or 12 h), Y-27632 (8 h) or cycloheximide (3, 9 or 24 h) at 37°C with 5% CO₂ for an indicated period of time.