Cytexpert version 2.2

T lymphocytes were isolated from the spleen of mice: spleen tissues were ground, and cells were resuspended in PBS. The percentages of T lymphocytes in the spleen to CD4+, and CD8+ T cells were tested in a flow cytometer. Direct labeled CD3-APC, CD4-FITC, and CD8-PE antibodies were added. The negative control group, CD3 positive control group, CD4 positive control group, and CD8 positive control group were set up at the same time. After placing for 30 min on ice, 400 µL PBS resuspension cells were added. The data were collected by flow cytometry (Beckman Coulter, Pasadena, CA, USA) and analyzed by CytExpert (version 2.2) (Beckman Coulter, Pasadena, CA, USA).

H9c2 were seeded (1 × 10⁶ cells) in 6 cm diameter dishes. The cells were pretreated with EGCG, L-164,041, or SB215505 for 30 min, and subsequently treated with IL-18 for 18 h. After treatment, the cells were harvested, washed, and resuspended in PBS. Cells were maintained on ice for 30 min. At least 10⁴ cells were collected and analyzed using a CytoFLEX LX Flow Cytometer (Beckman Coulter). The size of cells was determined using CytExpert version 2.2 (Beckman Coulter).