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. Pill pic chloroquine Chloroquine resistant malaria countries Chloroquine This drug is taken once a week, starting about 1 to 2 weeks before your trip and continuing for 4 weeks after. But chloroquine is rarely used anymore, because it no longer works. 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. 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. 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 Distinct effects of endosomal escape and inhibition of endosomal., Chloroquine Sensitizes Nasopharyngeal Carcinoma Cells but Not. Dosage of plaquenil 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. Chloroquine use in transfection. - Tissue and Cell Culture. Autophagy inhibition attenuates hyperoxaluria-induced.. Distinct effects of endosomal escape and inhibition of.. Hours after transfection reaction has been applied to cells, gently remove media and replace with 10% glycerol or DMSO for 2-3 minutes. Remove and feed with complete media various references. *1 hour before transfection, replace cDMEM with cDMEM + 25 mM chloroquine. FBS can be purchased already heat inactivated or it can be inactivated in the lab by heating to 56 ℃ for 30 minutes. 25 mM chloroquine diphosphate Dissolve 0.129 g of chloroquine diphosphate salt in 10 mL of sterile water. Filter sterilize through a 0.22 μm filter. Aliquot 50-100 μL and store at -20 ℃. Within 5-10’ of adding the chloroquine. 7. For 15cm plates, add 1.69ml of the transfection buffer to the DNA-CaCl2 solution by “bubbling”. To “bubble”, use two mechanical pipeters Pipet-AidTM, one with a 1ml pipette to bubble air into the DNA-CaCl2 solution and the other to slowly drip the transfection buffer into the conical tube.