Figure 7: GSH quantification using RT-based FACS in ferroptotic HT1080 cells and representative images.

(a) Histogram of ferroptotic HT1080 cells measured using RT on a flow cytometer. Cells were treated with erastin (10 μM) for 0 (pink), 6 (blue) and 24 h (turquoise). (b) Statistical analysis of ferroptotic HT1080 cells measured with FACS and LCMS. For FACS, each data point represents the mean value of >13,000 cells analysed from two independent experiments. Standard parametric unpaired t-test was used to analyse the data with P<0.0001 between groups. For LCMS, each data point represents the mean value of two independent experiments. Error bars represent s.d. (c) Representative images of HT1080 cells treated with 10 μM erastin. Scale bar, 10 μm. Despite significant morphology changes, GSH level did not change significantly over 3 min time span.