Fig. 7

Roles of PI3K/Akt and p53/p38 in phase conversion after Bu/Cy treatment. a Representative images of phosphorylated Akt⁺ cells among K15⁺ HFSCs in the bulge. b Representative images of phosphorylated p38⁺ cells among K15⁺ HFSCs in the bulge (immunofluorescence; scale bar = 50 μm). c Protein analysis after Bu and/or Cy treatment with PI3K inhibitor (20 µM; LY294002) comparing the concentrations of phosphorylated PI3K, phosphorylated and total Akt, cyclin D1, p53, and phosphorylated and total p38, with β-actin as a loading control (n = 4 biological samples, Supplementary Fig. 8a). d Protein analysis after Bu and/or Cy treatment with p38 inhibitor (10–20 µM; SB202190 or SB203580) comparing the concentrations of phosphorylated and total Akt, p53, phosphorylated and total p38, p21, and cleaved caspase-3, with β-actin as a loading control (n = 4 biological samples, Supplementary Fig. 8b). e Protein analysis after sequential Bu/Cy treatment with PI3K or p38 inhibitor showing the temporal changes of phosphorylated and total Akt, cyclin D1, p53, phosphorylated and total p38, p21, and cleaved PARP, with β-actin as a loading control. Temporal quantification plots of the concentrations of p53 per β-actin (n = 4 biological samples, Supplementary Fig. 9; Western blotting). Bu busulfan, Cy cyclophosphamide, Bu/Cy busulfan followed by cyclophosphamide, LY LY294002; SB SB202190 or SB203580, c-caspase-3 cleaved caspase-3, c-PARP cleaved PARP. Data are mean ± SEM. Source data are provided as a Source Data file. *p < 0.05 (vs. Con); **p < 0.01 (vs. Con, two-way analysis of variance with Dunnett’s multiple comparisons test)