Evaluating Cytotoxicity and Apoptosis in MCL Cells

These assays were performed as described previously [12 (link)]. In the cell viability assay, cells were seeded at 10,000 cells per well in 96-well plates and exposed to AZD4573, zelavespib, and tanespimycin for 72 h. Subsequently, cell lysis was conducted using the CellTiter-Glo Luminescent Cell Viability Assay Reagent, and luminescence was quantified employing the BioTek Synergy HTX Multi-mode microplate reader. For the apoptosis assay, Annexin V-binding was employed. MCL cells were treated separately with the vehicle, AZD4573, zelavespib, and tanespimycin, stained with Annexin-V and propidium iodide, and then subjected to flow cytometric analysis using the Novocyte Flow Cytometer to determine the percentages of Annexin-V positive cells. Data analysis was carried out using NovoExpress or FlowJo10, and each experiment was meticulously repeated at least three times to ensure the reliability of the results, consistent with the procedures outlined in the reference.