Analysis of T cell activation molecules in ECs by flow cytometry
BMMNCs were cultured in EGM-2-BulletKit medium and then incubated with 100 ng/ml LPS for 4 hours. Afterwards, to test the levels of T cell activation molecules in ECs, the cells were collected and stained with the previously mentioned EC markers and the following multicolour flow cytometry panel at room temperature for 15 minutes: anti-TLR4-BV786, anti-CCR5-BUV737, anti-CD80-BV650, and anti-CD86-BB515 (BD, Biosciences).