Fig 2: G-CSF induces mobilisation of HSCs into the peripheral blood.The frequency of circulating CD34+ and CD34+ CD133+ HSCs in the peripheral blood of participants with FA at baseline (day 1) or in response to G-CSF administration (day 5). Statistical comparisons are day 5 vs day 1 (baseline) using the sign test. *P = 0.031 (two-sided). Graph depicts mean ± standard error; n = 6 biologically independent samples. Source data are provided as a Source Data file.

Fig 3: QW24 decreases the self-renewal of colorectal CICs and migration of colorectal cancer cells. a-c, HCT116 cells were pretreated with indicated concentrations of QW24 for 4 h. The pretreated cells were cultured in unattached 96-well plates (1000 cells/well) to form the sphere as mentioned in the Methods, and the spheres images were taken after 1 week (a). The sphere number (b) and diameter (c) were measured. Scale bars, 100 µm. Data are presented as mean ± s.d. (n = 8); *, P < 0.05; ***, P < 0.001. d, QW24 pretreated HCT116 cells were seeded in unattached 96-well plates (1000 cells/well) and cultured for 1 week. The percentage of wells with spheres was determined and computed with limiting dilution analysis (LDA) to determine the sphere-initiating cell frequency. Frequency and probability estimates were computed using the ELDA software. n.s., Not statistically significant; ***, P < 0.001. e, HCT116 cells were treated with the indicated concentrations of PTC-209 or QW24 for 12 h, and cells were trypsinized, washed with PBS and stained with CD44-PE and CD133-APC antibodies. The stained cells were analyzed using flow cytometry. f, HCT116 cells were treated with 2 µM QW24 and the cells were imaged at the indicated time. Scale bars, 50 µm. g, HCT116, HT29 and CT26 cells were resuspended with 0, 1, 2, 4 µM QW24 in serum-free medium and seeded in transwell chambers for 12 h as descripted in Methods and the migrated cells were imaged and counted. Scale bars, 100 µm. Data are presented as mean ± s.d. (n = 3); **, P < 0.01; ***, P < 0.001

Fig 4: Examination of cancer stem cell markers in EAC-SCC cells. Cells were prepared on slides using Cytospin 4® (Thermo Fisher Scientific). EAC-SCC cells were tested with anti-CD44-PE, anti-CD133-APC, and anti-Oct-3 / 4 + Alexa Fluor® 555 anti-mouse IgG. DAPI (blue) was used to identify the location of cell nuclei for immunofluorescence staining. Fibroblasts were positive for CD44 and were not stained with CD133 or Oct3/4. In addition, ALP activity was observed in our EAC-SCC cells but not in fibroblasts. Scale bar, 50 µm.

Fig 5: RACK1 promotes self-renewal and chemoresistance of human liver CSCs and maintains murine ESC function. (A-N) 96 h after HuH7 cells were infected with lentivirus expressing non-targeting control (NC) shRNA or RACK1 shRNA (A-G), or after HuH7 single clone stably expressing FLAG-RACK1 and the mock control were generated (H-N), cells were subjected to the following assays: (A,H) Immunoblotting (IB) analysis of RACK1 expression in total cells. (B,I) Flow cytometric analysis of CD13 and CD133 expression in total cells. (C,J) Immunoblotting analysis of RACK1 expression in sorted CD13+ and CD13- subpopulations. (D,K) Sphere formation assays of sorted CD13+ subpopulation. mean±s.d. (n=3); *P<0.05, **P<0.01. (E,L) In vivo tumorigenicity experiments of sorted CD13+ subpopulation (5000 cells/site, 7 weeks, n=6). (F,M) Etoposide (Etop, 100 µM, 48 h)- or sorafenib (Sora, 50 µM, 24 h)-induced apoptosis of CD13+ subpopulation. mean±s.d. (n=3); Ctrl, control. (G,N) qRT-PCR analysis of sorted CD13+ subpopulation for the expression of the indicated drug-resistant relative genes. mean±s.d. (n=3). (O-Q) 96 h after murine ESCs were infected with lentivirus expressing non-targeting control shRNA or RACK1 shRNAs, cells were subjected to immunoblotting analysis for RACK1 expression (O), colony formation assays (mean±s.d., n=3; scale bar: 1 cm) (P), and alkaline phosphatase (AP) activity assays (mean±s.d., n=3) (Q).