Hydroxychloroquine sulfate hcq

Raw264.7, HT-29, HepG2, and HeLa cells were obtained from the American Type Culture Collection. Cells were maintained in DMEM (Gibco), 10% FBS (Gibco), 0.37% sodium bicarbonate (Sigma), and 4 mM l-glutamine (Hyclone) (full medium) at 37 °C and 5% CO₂. Earle’s balanced salt solution (EBSS; starvation medium) and hydroxychloroquine sulfate (HCQ) were obtained from Sigma. Bafilomycin A1 (Baf; LC laboratories) was used at 0.1 µM. The plasmid constructs encoding RFP-GFP-LC3B and GFP-LC3B used in this study have been described previously^{51 (link)}. For immunoblotting, polyclonal antibodies against p62 (Progen) were used at 1: 3,000 dilutions, polyclonal antibodies against LC3 (MBL International) were used at 1: 2,000 dilutions, polyclonal antibodies against V0a3 (Thermo Fisher) were used at 1: 2,000 dilutions, polyclonal antibodies against V1A1 (Thermo Fisher) were used at 1: 2,000 dilutions, and monoclonal antibody against Actin (Abcam) was used at 1: 10,000 dilutions. The fluorescent dye Hoechst 33342 (Invitrogen) was used at 1:500 and LysoTracker Red (LTR; Invitrogen) dye was used at 0.25 µM.

Atorvastatin and hydroxychloroquine sulfate (HCQ) were purchased from Sigma-Aldrich Co. LLC. (Shanghai, China) and SPH Zhongxi Pharmaceutical Co.,Ltd. (Shanghai, China), respectively. Atorvastatin was brought into suspension in normal saline (NS) at 0.286 mg/ml, and a 0.3-ml volume (equivalent to 2.6 mg/kg) was administered via oral gavage, once a day for six consecutive days a week. NS was orally administered as a vehicle control. The positive control group was orally administered HCQ (5 mg/kg/day). All mice (n = 30) were orally administered at 8 weeks of age for 6 consecutive days a week, and the surviving mice were killed at 19 weeks of age. The dose of Atorvastatin (2.6 mg/kg/day) is about approximately the dose given to human patients [19 (link), 20 (link)].