Transfections allow for transient expression of a gene of interest in a target cell line and can be useful for short term studies of protein function. We specifically use this protocol with Lenti-X 293T cells, a cell line optimized for production of lentiviral vectors. What is in plaquenil What is plaquenil therapy What is plaquenil Lung nodules plaquenil Chloroquine-containing media were removed and the cells were washed with phosphate buffered saline before they were infected with SARS-CoV 0.5 multiplicity of infection for 1 h in the absence of chloroquine. Virus was then removed and the cells were maintained in Opti-MEM Invitrogen for 16–18 h in the absence of chloroquine. Each Premo™ Autophagy Sensor Kit includes the BacMam LC3B-FP, a control BacMam LC3B G120A-FP mutation on the control BacMam LC3B prevents cleavage and subsequent lipidation during normal autophagy, thus protein localization should remain cytosolic and diffuse and chloroquine diphosphate to artificially induce autophagosome accumulation. Chloroquine is an antimalarial drug that has been used in humans for many years. In recent years, Chloroquine has been shown to inhibit autophagy and induce apoptosis in malignant cells and thus has been tested in various experimental model systems and in human clinical trials. Last Upload: June 10, 2016 Day 0: Seed Lenti-X 293T cells (this cell line is optimized for production of lentiviral vectors) Day 1 (pm): Transfect Cells Day 2 (am): 18h post transfection - Remove media, replace with fresh media Day 3 or more (am): Observe fluorescence, harvest cells, or perform your experiment *Pro-Tips* Different brands and lots of FBS can promote or inhibit transfection. This approach can be adapted for different cell lines and different transfection reagents. Chloroquine invitrogen Chloroquine for research Cell-culture tested InvivoGen, Premo™ Autophagy Sensor LC3B-GFP BacMam 2.0 Chloroquine for transduction hcmecPlaquenil drug interactions side effectsHydroxychloroquine sulfate 200 mg fc tabsWhat are the side effects of plaquenilChloroquine is a painless suicide C3556 Catalase from human erythrocytes ≥90% SDS-PAGE, buffered aqueous solution, ≥30,000 units/mg protein Catalase activates the decomposition of hydrogen peroxide, a reactive oxygen species, into water and oxygen. Autophagy Inhibitors - Autophagy Sigma-Aldrich. Chloroquine Is a Zinc Ionophore - PubMed Central PMC. Premo™ Autophagy Tandem Sensor RFP-GFP-LC3B Kit. Invitrogen Molecular Probes LC3B Antibody Kit for Autophagy 1 kit Life Sciences. Chloroquine diphosphate is also included - following treatment with this compound. Chloroquine prevented ductal carcinoma in situ xenografts’ outgrowth in athymic mice 37, 38 and inhibited N-methyl-N-nitrosurea-induced mammary carcinogenesis, suggesting chloroquine-based therapy as a possible agent in the prevention of initial premalignant lesions from progressing to breast cancer. Alexa Fluor R products are provided under an intellectual property license from Life Technologies Corporation. The purchase of this product conveys to the buyer the non-transferable right to use the purchased product and components of the product only in research conducted by the buyer whether the buyer is an academic or for-profit entity.