Supplementary Figure 7: RIPK1 inhibition, but not ROS neutralization, rescues the loss of mitochondria during ECM-detachment as a consequence of IDH2.

(a) H460 EV cells were grown in Att or Det for 24h treated with DMSO, Nec1 (10μM), MitoTEMPO (10μM), or M. Malate (2.5mM). Cells were then cytospun onto slides, fixed, and stained with Mito-Tracker Red (200nM) (red) and DAPI (blue). Representative images of each condition are shown (40x). Experiments were performed a minimum of three independent times with similar results. (b) Relative fluorescence measurements for each condition in (a) were determined using Fiji. For fluorescence calculations, n = 5 images. (c) H460 IDH1KO cells were grown in Att or Det for 24h treated with DMSO, Nec1 (10μM), MitoTEMPO (10μM), or M. Malate (2.5mM). Cells were then cytospun onto slides, fixed, and stained with Mito-Tracker Red (200nM) (red) and DAPI (blue). Representative images of each condition are shown (40x). Experiments were performed a minimum of three independent times with similar results. (d) Relative fluorescence measurements for each condition in (c) were determined using Fiji. For fluorescence calculations, n = 5 images. (e) H460 IDH2KO cells were grown in Att or Det for 24h treated with DMSO, Nec1 (10μM), MitoTEMPO (10μM), or M. Malate (2.5mM). Cells were then cytospun onto slides, fixed, and stained with Mito-Tracker Red (200nM) (red) and DAPI (blue). Representative images of each condition are shown (40x). Experiments were performed a minimum of three independent times with similar results. (f) Relative fluorescence measurements for each condition in (e) were determined using Fiji. For fluorescence calculations, n = 5 images. All results are presented as means +/- SEM and statistical significance was calculated using a Student’s two tailed t-test. p<0.05 is statistically significant. N.S., not significant. Scale bars, 100 μm.