2.10 Flow cytometric analysis
A549 cells or MLE-12 cells were seeded in a twelve-well plate with a density of 2×105 cells/well in complete growth medium. Following with or without 5 mM lysine treatment, the cells were exposed to PQ in a dose dependent manner, as indicated, for 24h, washed twice with PBS and incubated with 2 µM Fluo-3/AM (Beyotime, S1056) diluted in Hanks’ Balanced Salt Solution (Beyotime, C0218) for 30 min at 37°C in the dark. The stained cells were then dissociated with trypsin, resuspended with PBS and collected to detect the presence of [Ca2+]i using a FACScan flow cytometer (BD LSRFortessa™, NJ, USA).