Flow cytometry
For the apoptosis assay,
cells
were
collected
after treatment, and an
Annexin
V/PI detection kit
(BD
Biosciences) was used to detect apoptosis by flow cytometry according to
the protocol described. For the ROS assay,
cells
were incubated with the probe DCFH-DA at a concentration of 10 μM after
the
different
treatments for 30 min at 37 °C, and then
flow
cytometry was used for fluorescence intensity detection. For
the
mitochondrial ROS assay, cells subjected to the different treatments
were incubated with the probe mitoSOX at a concentration of
5
μM for 10 min
at
37
°C,
and then flow cytometry was used for fluorescence intensity detection.
For the plasma membrane potential assay, cells were incubated in
KRH
(Krebs-Ringer-HEPES) buffer containing 100 nM DiBAC4(3) for 20 min at
room temperature before flow cytometry detection. For the
mitochondrial
membrane potential (MMP) assay, TMRM was also detected by flow
cytometry.