Countess c10281 automated cell counter

CAFs (3.0 × 10³ cells) in 50 µL of FGM2 were seeded into the upper chamber of a Transwell plate (Corning HTS Transwell 96 wells, #CLS3388, Corning Inc.), and 200 µL of FGM2 was added to the lower well. After 24 h, CAFs were transfected with siRNAs as described above and incubated for 48 h at 37 °C. Medium in the upper chamber was subsequently replaced with 50 µL of AIM V serum-free medium (Thermo Fisher Scientific) with 10% human serum (Biowest, Nuaillé, France) containing 1 × 10⁵ CD8+ T lymphocytes. The medium in the lower well was replaced with 200 µL of culture medium with 10% human serum and recombinant human CXCL10 (1 ng/mL, Shenandoah Biotechnology, Warminster, PA, USA). After incubation for an additional 24 h, cells in the lower chamber were counted using a Countess C10281 automated cell counter (Thermo Fisher Scientific).