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. Chloroquine sensitive malaria areas Deubiquitinating enzyme chloroquine Does plaquenil cause leg pain Transfection of SW480 human colorectal cancer cells was highly efficient with transfection efficiencies up to 100 % using optimized conditions of K2 Transfection System Biontex. Transfection efficiency and expression were higher than that of a commercially available transfection reagent x, commonly used in our lab. In contrast, in the present study our results indicated that oxalate-induced mitochondrial edema and depolarization, as well as the oxidative injury of renal tubular cells, were markedly attenuated by chloroquine pretreatment but accelerated by rapamycin pretreatment, suggesting that autophagy activation has detrimental effects on oxalate-induced mitochondrial damage and oxidative injury in renal tubular epithelial cells. Chloroquine is an inhibitor of the lysosomal degradation of the DNA which is taken up by the cells, so as leelee said, transfection should have been successful, albeit at a slightly lower level than if you would have added the chloroquine. In my normal transfections, I never add chloroquine and get high efficiencies nevertheless. 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 pretreatment before transfection BRIEF COMMUNICATION Chloroquine and amphipathic peptide., Autophagy inhibition attenuates hyperoxaluria-induced. Hydroxychloroquine lose eye sight In the current study, we observed that pretreatment of COS7 and HEK293 cells with these lysosomotropic agents, chloroquine 100 μM and ammonium chloride 10 mM, for 4 hours prior to electrotransfection insignificantly altered eTE Fig 7a and 7b. The pretreatment was required because the lysosomotropic agents need to be in lysosomes before. Distinct effects of endosomal escape and inhibition of.. Chloroquine use in transfection. - Tissue and Cell Culture. Autophagy protects against palmitate-induced apoptosis in hepatocytes.. Pretreatment with chloroquine before radiation increases the percentage of cells with morphological signs of apoptosis condensed and fragmented nuclei in radiation-sensitive NPC cell lines CNE-2 and HNE-1, but not in the radioresistant cell line C666-1 and the immortalized nasoepithelial cell line NP69. Hemozoin collects in the digestive vacuole as insoluble crystals. Chloroquine enters the red blood cell, inhabiting parasite cell, and digestive vacuole by simple diffusion. Chloroquine then becomes protonated to CQ2+, as the digestive vacuole is known to be acidic pH 4. 7; chloroquine then cannot leave by diffusion. Dilute 100 mM chloroquine diphosphate 11,000 directly into the medium either before or after the addition of the calcium phosphate–DNA coprecipitate to the cells Step 3.