Flou-3-pentaacetoxymethyl (Fluo-3/AM; Beyotime) was used for intracellular Ca2+ testing. K562 cells from each group were collected and resuspended with 1 μmol/L Fluo-3/AM diluted in 1 mL D-Hank's balanced salt solution (Solarbio, Beijing, China) for 30 min. Then, the cells were incubated for another 30 min and analyzed by a C6 flow cytometer (BD Biosciences).
A 5,5′,6,6′-tetrachloro-1,1′,3,3′-tetraethyl-imidacarbocyanine (JC-1) kit (Beyotime) was used for MMP measurement. K562 cells at 24 h after coculturing were stained with 0.5 mL JC-1 solution. All samples were incubated for 20 min and detected by a C6 flow cytometer.
2′,7′-Dichlorofluorescein diacetate (DCFH-DA, Beyotime) was used for ROS determination. Twenty-four hours after coculturing, K562 cells were stained with 10 μL of DCFH-DA for 20 min and resuspended in D-Hank's solution, and 2′,7′-Dichlorofluorescein (DCF) fluorescence was also detected by a C6 flow cytometer.
A 5,5′,6,6′-tetrachloro-1,1′,3,3′-tetraethyl-imidacarbocyanine (JC-1) kit (Beyotime) was used for MMP measurement. K562 cells at 24 h after coculturing were stained with 0.5 mL JC-1 solution. All samples were incubated for 20 min and detected by a C6 flow cytometer.
2′,7′-Dichlorofluorescein diacetate (DCFH-DA, Beyotime) was used for ROS determination. Twenty-four hours after coculturing, K562 cells were stained with 10 μL of DCFH-DA for 20 min and resuspended in D-Hank's solution, and 2′,7′-Dichlorofluorescein (DCF) fluorescence was also detected by a C6 flow cytometer.