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. How does plaquenil work for sjogren& 39 Leflunomide hydroxychloroquine combination Treatments may modify cell membrane structure to enhance uptake of DNA. Chloroquine, which is generally applied to cells with the DNA, appears to enhance transfection efficiency by inhibiting DNA degradation by lysosomes 15. Sodium butyrate treatment following transfection has been reported to result in higher levels of transient expression 16. The total volume of transfection complex medium, DNA and ViaFect™ Transfection Reagent to add per well of a 96-well plate is 5–10μl. The following protocol is a guideline for transfecting approximately 10–20 wells, depending on the volume of ViaFect™ Transfection ReagentDNA mixture used. For additional wells, scale volumes accordingly. A pH-responsive peptide fragment modelled on the influenza virus haemagglutinin INF7-SGSC can promote the transfectional activity of polyL-lysine pLL/DNA complexes against 293 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 dna transfection Transfection Methods Overview - Bio-Rad, Transfection Guide Overview of Transfection Methods Promega Hydroxychloroquine hair loss Chloroquine treatment of rodent cells during the first hours of polyoma DNA transfection increase the fraction of cells expressing viral functions. The effect has been observed after DNA absorption using both the DEAE-dextran and calcium phosphate coprecipitation methods. High efficiency polyoma DNA transfection of chloroquine.. Chloroquine and amphipathic peptide helices show synergistic.. Helper Dependent Protocol - Stanford University. Add DNA/CaCl2-solution to 2x HBS drop wise, while blowing air through HBS with pasteur pipette. • Incubate the mixture for 10 to 20min at room temperature. • Remove old medium from the cells. • Add 10ml medium including 25µM Chloroquine the Chloroquine is used in this step only. Chloroquine has been used to increase transfection efficiency for calcium phosphate, DEAE-dextran, and some liposome-mediated gene delivery 63,65–68. However, effects of lysosomotropic agents on polymer and some lipid based methods of gene delivery are still controversial in the literature. Transfection-mediated toxicity usually leads to changes in gene expression, even in the absence of any visual toxicity. Therefore, it is important to choose a transfection method that can mitigate both observed as well as non-apparent cellular toxicity. Cationic lipids and polymers also form complexes with DNA through electrostatic interactions.