Jurkat cells

HeLa cells, L929 cells, U251 cells, and Jurkat cells were purchased from the Korean cell line bank (Seoul, Republic of Korea) and used for the following experiments. HeLa (cervical cancer cell), L929 (murine skin fibroblasts), and U251 (glioma cell) cells were cultured in DMEM (Welgene, Gyeongsan-si, Republic of Korea), supplemented with 1% antibiotic antimycotic solution, 100× (Corning, Manassas, VA, USA) and 10% fetal bovine serum (Welgene, Gyeongsan-si, Republic of Korea). Jurkat (T-Cell Lymphocyte leukemia) cells were cultured in RPMI (Welgene, Gyeongsan-si, Republic of Korea), supplemented with 10% FBS and 1% Abs. All cells above were cultured in a 5% CO₂ chamber at 37 °C. C2C12 (mouse myoblasts) cells were a gift from Mi-Ock Lee at Seoul National University in Seoul, Republic of Korea. C2C12 cells were cultured in DMEM supplemented with 10% FBS and 1% antibiotic antimycotic solution, 100× (Biowest, Riverside, MO, USA). For the induction of differentiation, fully confluent C2C12 cells were cultured in DMEM supplemented with 2% horse serum (Sigma-Aldrich, St. Louis, MO, USA) for 3 days.

Jurkat cells were purchased from the Korea Cell Line Bank (Seoul, Republic of Korea) and maintained in RPMI-1640 medium containing 10% FBS, streptomycin sulphate, penicillin, HEPES, and sodium bicarbonate in a 5% CO₂ atmosphere at 37 °C. Jurkat cells were stimulated with 1 μg/ml of plate-bound anti-CD3 mAb (clone HIT3a, BD Bioscience) and then incubated with KITS 1–001 (10, 50, or 100 μM) for 30 min. KIST 1–001 were dissolved in DMSO and added to the culture media in serial dilution (the final concentration of DMSO in all experiments did not exceed 0.1%).