Mission turbogfp

The MCF7 human breast cancer cell line was obtained from the American Type Culture Collection (ATCC) and cultured using RPMI medium (GIBCO, Invitrogen—Carlsbad, CA, USA) containing 10% FBS (GIBCO, Invitrogen) and 1% Pen/Strep (GIBCO, Invitrogen), at 37 °C with 5% CO₂ in a humidified atmosphere. For constitutive GFP labeling, cells were transduced using a lentiviral-driven GFP construct (Mission TurboGFP, SHC003V, Sigma—San Luis, Misouri, USA) following the manufacturer’s instructions. GFP-positive cells were selected 72 h post-infection, adding 10 µg/mL puromycin to the culture media.

Lentiviral particles expressing a green fluorescence protein (GFP) (MISSION TurboGFP, Sigma-Aldrich, St. Louis, MO) were used to infect cell cultures with the manufacturer’s recommended protocol. Briefly, cell cultures in 0.5 ml culture medium in each well of a 6-well plate were infected with 10 μl (1 × 10⁷ TU) of viral particles. Polybrene (Santa Cruz Biotechnology, Dallas, TX) was used at 8 μg/ml to enhance infection efficiency. After 4 h of incubation at 37°C, 1.5 ml fresh medium was added, and the cultures were maintained for 4 weeks with daily fluorescence microscope inspection.