For FACS analysis, cells were first dissociated with 0.05% Trypsin in 0.2% EDTA and PBS.
For DNA fragmentation analysis, cells were treated with 1 μg/mL for 12 h.
For FACS analysis, cells were collected by using TrypLE Express (Gibco) and measured by a flow cytometer (BD LSRFortesa).
For protein expression analysis, cells were harvested at 48 hours.
For cytochrome c release and mitochondrial protein analysis, cells were fractionated into mitochondrial and cytoplasmic fractions.
For immunofluorescence analysis, cells were grown on glass coverslips inside 6-well plates.
For flow cytometric analysis, cells were fixed and permeabilized with BD Cytofix/Cytoperm (Becton Dickinson).
Cell cycle analysis: Cells were resuspended in 200 µl 7-AAD binding buffer (BD Pharmingen).
For this analysis, cells expressing Cherry-tTA-ER at equivalent intensity levels were selected for imaging.
Prior to analysis, cells were filtered through a cell strainer cap (30 µm) to remove debris.
For cell cycle analysis, cells transfected in Greiner 12-well culture dishes (cat. no.
