Measurement of ROS levels
After treatment with different concentrations of icariin for 24 h, the
cells were harvested and incubated with DCFH-DA for 20 min at 37ºC. The
ROS levels of treated cells were measured via flow cytometry (FCM). For
inhibition experiments, cells were treated with an antioxidant (NAC, 2
mM) for 1 h prior to icariin exposure.
Mitochondrial membrane
potential (ΔΨm) assay
Rh123 was exploited to measure the changes
in the ΔΨm of cells
via FCM. Cells were treated with different concentrations of icariin and
then incubated with Rh123 (5 μg/mL) at
37ºC for 30 min. Finally, the
ΔΨm of the treated cells was measured by FCM.