Lipofectamine

Female C57BL/6 mice were purchased from The Jackson Laboratory or Charles River Laboratories. F5 mice that are transgenic (Tg) for nucleoprotein of influenza virus A/NT/60/68 (³⁶⁶ASNENMDAM³⁷⁴;NP68)-specific, H-2D^b–restricted T-cell receptor were obtained from Taconic Farms (Hudson, NY). All mice were housed in microisolator cages in pathogen-free conditions. Mice used for the in vivo antitumor studies were 16 to 18 weeks old at the start of study. Animal care was in compliance with The Guide for Care and Use of Laboratory Animals (National Research Council).
The MB49 parental cell line (murine transitional cell carcinoma) was kindly provided by Dr. Peter Pinto (Urologic Oncology Branch, CCR, NCI, NIH). Cells were grown, batch frozen and used in the experiments described. The MB49 LucSH⁺ cells (MB49^luc) were generated within our laboratory. All MB49 lines were grown in Dulbecco’s Modified Eagle Medium (DMEM) with 10% heat-inactivated fetal bovine serum supplemented with 1 mM nonessential amino acids, 1 mM sodium pyruvate, 2 mM glutamine, and penicillin/streptomycin (100 U/mL). MB49^luc growth medium also contained Zeocin (200 µg/ml). MB49^luc are parental MB49 cells transfected with a pSELECT-zeo-LucSh plasmid using Lipofectamine (InvivoGen, San Diego, CA) for luciferase expression detected by in vivo imaging.