Extended Data Fig. 4: ER Ca2+ storage contributes to ferroptosis.
From: CGI1746 targets σ1R to modulate ferroptosis through mitochondria-associated membranes
a-b. Compound CGI1746 did not protect U937 cells from apoptosis (a) and necroptosis (b). c. Cell viability of NCI-H1299 cells treated by RSL3 for 9 hours with or without BAPTA-AM (5 μM) pre-treatment. d. The morphology of NCI-H1299 cells treated with RSL3 (2 μM) for 8 hours following BAPTA-AM pre-treatment (5 μM). Scale bar, 50 µm. e-f. Cell viability of PC-3 (e) and MEF (f) cells treated by RSL3 for 10 (e) and 12 (f) hours respectively with or without BAPTA-AM (10 μM) pre-treatment. g. MEF cells were treated by erastin for 12 hours with or without BAPTA-AM (10 μM) pre-treatment for cell death analysis. h. Cell survival rates of PC-3 cells incubated with RSL3 for 24 hours in the presence or absence of BAPTA-AM (10 μM). i. Cell death at 48 hours in NCI-H1299 cells treated by erastin after pre-treatment with or without BAPTA-AM (10 μM). j. The effect of BAPTA-AM (10 μM) on ML210 induced ferroptosis in NCI-H1299 cells for 6 hours. k. Cell death in MDA-MB-468 cells treated by tBOOH plus Z-VAD (20 μM) for 6 hours with or without BAPTA-AM (10 μM) pre-treatment. l. The capacity of antioxidant activity of BAPTA-AM (5 μM) in DPPH assay. m-o. The effect of EGTA-AM (20 μM) on ferroptosis in MDA-MB-468 cells (m), NCI-H1299 cells (n), MEF cells (o). p. The mRNA levels of SERCA1, SERCA2 and SERCA3 by quantitative real-time RT-PCR in NCI-H1299 cells. Two pairs of primers were used. Viability data represent mean ± s.d. of n = 3 replicates from one representative of three independent experiments. For bar graphs k and p, data represent mean ± s.d. of n = 9 (k) and 3 (p) replicates from one representative of three independent experiments. Micrograph data show one representative out of three replicates from one representative of three independent experiments. One-way ANOVA (GraphPad Prism 9.5.1), ****P < 0.0001 and ns, not significant.
