Fig. 5

Aberrancies in cell-cycle status and apoptosis in cultured KRAS (G13C) iPSC-HPCs. a Flow cytometry-based cell-cycle analysis of control (C2-1) and mutant (R2-1) iPSC-HPCs expanded under either the SF (Selective) or STF3 (Standard) condition. Representative plots are shown with the estimated percent values of each cell-phase. G1-phase, lower left; S-phase, top; G2/M-phases, lower-right. b Quantitative comparison between C2-1 and R2-1 HPCs (cell percentage) in S-phase. Mean ± SD values are shown (independent experimental replicates: n = 3). c Cell viability analysis of C2-1 and R2-1 HPCs expanded with indicated culture conditions. With the samples processed for the cell cycle assessment, the events appearing as a distinct population characterized by a reduced DNA content (low fluorescent intensity with the FxCycle™ Violet Stain) were visualized and quantified as sub-G0/G1 cells using flow cytometry. Mean ± SD values are shown (independent experimental replicates: n = 3, Selective; n = 4, Standard). NS, not significant. d Detailed cell death analysis of C2-1 and R2-1 HPCs cultured in the standard condition. Representative images are shown in two-dimension plots with Annexin-V and 7AAD used as cell-death indicators. The estimated percentage for the events contained in each quadrant is indicated. e Quantitative comparison between C2-1 and R2-1 iPSC-HPCs in percentages of the cells in each quadrant. Mean ± SD values are shown (independent experimental replicates: n = 3). All statistical analyses are based on an unpaired t test. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. NS, not significant