Fig. 3: Loperamide, pimozide- or STF-62247-induced cell death does not primarily involve apoptosis, ferroptosis, or necroptosis.

a, c, d MZ-54 cells were pretreated for 1 h with 20 µM zVAD.fmk (a), 5 µM Fer-1 (c) or 30 µM Nec-1s (d) followed by treatment with 17.5 µM loperamide, 15 µM pimozide, 40 µM STF-62247, 20 µM IM/100 µM TIC, 25 µM ABT-737/100 µM etoposide, 500 nM RSL3 or 1 ng/mL TNF\({\alpha}\) + 0.5 µM BV6 for 48 h. Cell death was assessed by measuring the PI uptake as fraction of total nuclei determined by Hoechst counterstaining using high-content fluorescence microscopy. HT-29 cells served as positive control for induction of necroptotic cell death. b MZ-54 cells were treated with 3 µM STS, 15 µM loperamide, 15 µM pimozide, 40 µM STF-62247, or 20 µM IM/100 µM TIC for the indicated time points. Caspase-3 activity was determined by quantifying alterations in Ac-DEVD-AMC fluorescence. Mean and SEM of 3−4 independent experiments performed in triplicate are shown. *p < 0.05, **p < 0.01, ***p < 0.001. UT untreated, LOP loperamide, PIMO pimozide, STF STF-62247, IM imipramine hydrochloride, TIC ticlopidine, STS staurosporine