BPAE imaging using CellROX® Deep Red Reagent (Cat. No. C10422)

Bovine pulmonary artery endothelial (BPAE) cells were plated in a 96-well plate. BPAE cells were treated with or without 100 µM menadione for 1hour at 37°C. 100 µM superoxide scavenger, MnTBAP, was added to some of the control and menadione-treated wells for the last 30 minutes of incubation. The cells were then stained with 5 µM CellROX® Deep Red Reagent and Hoechst 33324 by adding the probe to the complete medium and incubating the cells at 37°C for 30 minutes. The cells were then washed with PBS and imaged.