Caspase-Glo® 3/7 assays. 22Rv1 cells were seeded at 10,000 cells/well of white 96 well plates (Corning #3917) 12-24h before treatment with test compounds with and without inhibitors or an equivalent volume of DMSO for 24h. 100µL/well of Caspase-Glo 3/7 Reagent (Promega) was added and the plate shaken at 500 rpm for 30 s. The plate was incubated for 2h and luminescence measured using a PHERAstar FS plate reader (BMG Labtech). Clonogenic assay. 22Rv1 cells were treated with 10nM SIM1, cis-SIM1, MT1, MZ1 and ARV-711 for 24h. The next day, cells were trypsinised and counted. 500 cells were re-plated and allowed to grow at 37ºC and 5% CO2 for 20 days. After 20 days incubation, the cells were fixed with ice-cold 100% (v/v) methanol for 30 min at 4ºC. Afterwards, methanol was removed, and the cells were stained with 500µl 0.1% crystal-violet dye (in MeOH) for 30 min at room temperature. Following incubation, the cells were washed with dH2O and left to dry overnight. Plates were scanned on an Epson Perfection V800 Photo scanner. And image analysis was done in ImageJ software v. 1.52n. Plating efficiency (PE) was calculated by counting colonies for each treatment condition and dividing the average by number of cells plated. Survival fraction was determined by diving PE of treated cells by PE of untreated cells, multiplied by 100 . Bar graphs were generated using GraphPad Prism software. Two independent experiments were performed. Flow Cytometry. MV4;11 cells were counted on a Countess 3 Automated Cell Counter (Thermo Fisher, UK) with the addition of trypan blue. Cells (1x10 6 ) were aliquoted, spun down and resuspended in RPMI media containing test compounds at indicated concentrations. Additionally, QVD OPh (Sigma-Aldrich, UK) and Necrostatin-1 (Sigma-Aldrich) were added to the SIM1 treatment at 20µM final concentration. Treated cells were left to incubate for 24h at 37°C and 5% CO2. On the following day, the cells were collected in a Falcon tube and spun down at 500g for 5 min. Supernatant was aspirated and cells were washed once in 1mL FACS buffer (PBS, 5% FBS, 0.05 % NaN) and afterwards resuspended in 100µL of the same buffer containing Apotracker Green (Biolegend, UK) and DAPI (Sigma-Aldrich) at final concentration of 400nM and 1µg/mL, respectively. Cells were incubated for 20 min on ice and afterwards washed in 1mL of FACS buffer and finally resuspended in 500µL of the same buffer. Measurements were done on BD FACS Canto II flow cytometer (Flow Cytometry and Cell sorting facility, University of Dundee, UK) using blue (ex: 488 nm; em: 530±30 nm) and violet (ex: 405 nm; em: 450±50 nm) laser for detection of FITC and DAPI, respectively. Data were analysed on FlowJo™ 10.7.1. Software and GraphPad Prism. Gating strategy is detailed in Supplementary Figure .