Colon Cancer Cell Transfection Protocol

Glutamine, DMEM, trypsin-EDTA, and penicillin/streptomycin were obtained from GE Healthcare HyClone (Logan, UT, United States). Fetal bovine serum, enzymes, 3-aminopropyltriethoxy silane (TESPA), L-15 and chemicals were from Sigma (St. Louis, MO, United States). DiI-C₁₂, DiD-C₁₈, Lipofectamine 2000 and Gibco Opti-MEM were from Thermo Fisher Scientific (Waltham, MA, United States). High performance, 1.5H, coverslips were from Marienfeld (Lauda-Königshofen, Germany). The colon cancer cell line HT29 was a kind gift of Dr. A. Blokzijl, Uppsala University, Sweden and SW480 cells were from ATCC (Manassas, VA, United States). CD3ζ-EYFP, mouse CD3ζ fused to EYFP via a six aa linker expressed in the pBJ1-Neo plasmid under the CMV-promotor, was from Mark Davis, Stanford University School of Medicine, United States (Krummel et al., 2000 (link)). Lck-EGFP, a fusion protein of mouse Lck and EGFP via a six aa linker expressed in the pcDNA3 under the CMV-promotor, was from Tony Magee, Imperial College London, United Kingdom (Janes et al., 1999 (link)). Borosilicate glass (1B100F-4) was from World Precision instruments (Sarasota, FL, United States).