Notes: HUVECs, prior to passage 7, were transfected using the ViaFect™ Transfection Reagent (no further details provided). (4689)

Notes: The authors used the ViaFect™ Transfection Reagent to transfect various expression vectors into HEK 293 cells. In one study for immunoprecipitation and Western blotting, cells were grown to 70% confluence on 10-cm plate and transfected with 7µg of DNA. (4680)

Notes: Ghrelin receptor expression vectors were transfected into HEK 293 cells with the aid of the ViaFect™ Transfection Reagent (2.5µg plasmid DNA into 1 × 10⁶ cells growing in a collagen-coated 10cm dish). (4686)

Notes: The effect of overexpression of MAP2K1 (D67N) was assessed in the Calu-1 human cells through transfection with ViaFect™ Transfection Reagent (details not provided). The effect of 72 hour cisplatin exposure on 13 different human lung cell lines was assessed with the CellTiter 96® AQueous One Solution Assay. All cells were authenticated through STR analysis using the GenePrint® 10 System. (4682)

Notes: A GFP-Izumo1 expression vector was transfected into HEK 293 cells using ViaFect™ Transfection Reagent prior to western analysis. (4678)

Notes: HepG2/C3A cells were reverse transfected in collagen-coated 6 well plates with TRF2 constructs using ViaFect™ Transfection Reagent. Transfected cells were transferred to collagen-coated glass coverslips and used for immunofluorescent studies. (4679)

Notes: The 3’ UTR of the human SNCA message was subcloned into the psiCHECK™-2 Vector and used to screen siRNAs for interaction. Results were determined through the use of the Dual-Luciferase® Reporter Assay System. Identified siRNA duplexes were transfected into human fibroblasts possessing SNCA locus triplication seeded into 6-well plates at 1 × 10⁵ cell/well using 10nM final concentration utilizing ViaFect™ Transfection Reagent. Twenty-fours hours after transfection, RNA was extracted and analyzed. [The authors judged transfection efficiency for ViaFect and these cells by transfecting a GFP expression plasmid and visualizing the GFP positive cells (results in Supplementary Figure S4)]. (4681)

Notes: The ViaFect™ Transfection Reagent was used to perform transfections into HeLa cells and Vero cells, and ultimately used for fluorescent microscopy (details of the transfections were not provided). HeLa cells were the cell line for a split-GFP complementation assay with subsequent fluorescent microscopy. Vero cells were transfected with MxA-expression plasmid prior to viral infection to investigate localization of MxA and viral ribonucleoprotein. Firefly experimental and Renilla control vectors were used in a minimal replicon reconstitution assay. Successful replication of the Firefly construct was read via the Dual-Luciferase® Reporter System. (4676)

Notes: The effect of an miRNA was assessed in the NKN1, NKN1-EBV, NUGC3-EBV, AGS and AGS-EBV cell lines transfected with a dual-reporter vector containing the BID 3’ UTR sequence. The reporter vector was transfected using the ViaFect™ Transfection Reagent. After transfection with miRNA, the reporter levels were determined with the Dual-Glo® Luciferase Assay System. (4684)

Notes: Transfection of religated BK polyomavirus (BKPyV) genomic DNA into primary renal proximal tubule epithelial cells (RPTECs) was through the use of the ViaFect™ Transfection Reagent. Cells were transfected in 6-well plates at 90-95% confluency using a 3:1 ViaFect™ Reagent:DNA ratio. (4683)

Notes: GFP-based reporter plasmids were transfected into mouse male germline stem cells (GSCs) and mouse endothelial fibroblast cells (MEFs) using ViaFect™ Transfection Reagent (details not provided). (4687)

Notes: A bicistronic firefly/Renilla construct was engineered and transfected into 1.25 × 10⁵ MCF7 cells/well of a 12-well plate using the ViaFect™ Transfection Reagent. Reporter activities were measured with the Dual-Luciferase® Reporter Assay System and read on a GloMax® 20/20 Luminometer. (4688)

Notes: Human TE671 cells were tested for expression and glycosylation of two proteins, A1AT and GM-CSF. Initial work focused on comparison of TurboFect Transfection Reagent (Life Technologies) and AppliFect LowTox (AppliChem) for expression of A1AT. AppliFect proved superior for expression of the protein and maximum expression achieved through optimization. Both TurboFect and AppliFect were examined for expression of GM-CSF and low transfection efficiencies were obtained despite optimization of conditions and the researchers turned to ViaFect™ Transfection Reagent for improvement. The standard protocol for ViaFect™ Reagent produced results similar to optimized AppliFect but through optimization, the ViaFect™ Reagent were able to produce much higher protein yields (8 × 10⁴ cells; 0.5µg DNA/2µl ViaFect™ Reagent in 24-well plates). Enough protein was produced from transiently transfected cells, scaled to a 10cm dish, to allow development of an N-glycan profile for the recombinant GM-CSF. Interestingly, the authors went back to examine whether ViaFect™ could improve A1AT expression but did not find a difference large enough to change A1AT expression protocols. (4672)

Notes: The putative promoter and deletion mutants of the proposed promoter of the RBHG gene were cloned into the pGL3-Basic Vector for reporter assay investigation. Reporter plasmids and the pRL-TK Control Vector were cotransfected into HepG2 cells with the ViaFect™ Transfection Reagent (transfection details not provided). Forty-eight hours post-transfection, reporter activity was measured with the Dual-Glo® Luciferase Assay System using a GloMax® Instrument. (4685)

Notes: Epiprofin (Epfn) was expressed in HaCaT cells from various HaloTag® Fusion vectors with either full-length or deletion mutants of the CMV promoter to provide different levels of Epfn expression. (The authors do not state whether the N-terminal or C-terminal HaloTag® Fusion vectors were used). HaCaT 72hr proliferation was highest when using the CMVd3 promoter (e.g., pFC17A or pFN24A) for the lowest expression level. Reporter assays were performed on in the HaCaT cells as well and were measured with the Dual-Luciferase® Reporter Assay System on a GloMax® Instrument. Transfection studies performed with primary keratinocytes utilized the ViaFect™ Transfection Reagent using the reverse transfection method. (4690)