Fig. 1

Melphalan treatment induces cell death and apoptosis in hiPSC-CMs. a Representative images of hiPSC-CMs treated with melphalan for 5 days. Scale bar, 40 μm. b Measurement of cell viability by CellTiter-Blue Viability assay in hiPSC-CMs treated with melphalan for 3 and 5 days, respectively (n = 4). c Quantification of ATP content/well which indirectly showed viability by CellTiter-Glo 3D Viability assay in hiPSC-CMs treated with melphalan for 3 days (n = 4). d, e Representative images and quantification of cell apoptosis in hiPSC-CMs upon melphalan treatment for 24 h by CellEvent Caspase-3/7 Green Detection reagent and Hoechst staining (n = 4). Cells positive for activated caspase-3/7 emitted bright green nuclear fluorescence. Scale bar, 50 μm. f qRT-PCR analysis showing relative gene expression levels of apoptosis-related genes BCL2 and BAX in hiPSC-CMs treated with melphalan for 3 days (n = 3). The viability and relative MFI were normalized by the average values of no melphalan group. Comparisons were conducted between each treatment group and no melphalan group via one-way ANOVA test. **P value < 0.01; ***P value < 0.001; ****P value < 0.0001