Fig 2: Screening of miRNAs Regulating CD133 Expression and miRNA validation by RT-PCR. (A–L) represent the expression level of hsa-miR-29a-3p, hsa-miR-29b-3p, hsa-miR-200c-3p, hsa-miR-4423-5p, hsa-miR-335-3p, hsa-miR-142-3p, hsa-miR-142-5p, hsa-miR-22-3p, hsa-miR-30a-5p, hsa-miR-30e-5p, hsa-miR-377-3p and hsa-miR-4739 between CD133+HUCB-MNC cells and CD133−HUCB-MNC cells at 0 and 6 Gy.

Fig 3: CD133+HUCB-MNC cells were more radioresistant compared with CD133−HUCB-MNC cells. (A) Represents the survival curves for each cell type. (B) Representative hematopoietic colony formation culture images of CD133− cells (upper) and CD133+ cells (lower) at 8 Gy radiation dose. Scale bar = 50 μm. (C) Represents the growth rate of CD133− cells and CD133+ cells at 4 Gy, 6 Gy and 8 Gy radiation dose. (D,E) Represent the apoptosis proportion in the two type’s cells. (F,G) Represent the the effect of CD133 on the cell cycle.

Fig 4: OSM and Sal induce expression of LCSC differentiation-related markers. (A) Fraction of LCSCs expressing CD133 after treatment with OSM, Sal, or both according to flow cytometric analysis. (B) Real-time quantitative reverse transcription polymerase chain reaction analysis of NANOG, OCT4 and c-MYC mRNA expression in LCSCs treated with OSM, Sal, or both. Detection of (C) AFP and (D) ALB levels in the culture supernatant of LCSCs treated with OSM, Sal, or both by enzyme linked immunosorbent assay. OSM (10 ng/ml), Sal (1 µM), or both (10 ng/ml OSM + 1 µM Sal) were used in the experiments. Cells treated with 1 µg/ml 5-FU served as the positive controls, and culture medium was used as a negative control. Data represent the mean ± standard deviation. *P<0.05, compared with the control; #P<0.05, compared with 5-FU, ∆P<0.05, compared with the combination (OSM + Sal). OSM, oncostatin M; Sal, salinomycin; LCSC, liver cancer stem cell; AFP, α-fetoprotein; ALB, albumin; 5-FU, 5-fluorouracil.

Fig 5: Anisomycin inhibits HuOCSC invasion and angiogenesis in vitro. (A) A Transwell invasion assay was used to investigate the invasive properties of CD44+/CD133+ HuOCSCs following anisomycin treatment (n=4). Magnification, ×100. (B) Representative images and (C) quantification of the ability of HUVECs to form 3-dimensional tubular structures in Matrigel following anisomycin treatment (n=4). Magnification, ×100. (D) Quantification and (E) representative images of the growth and development of intersegmental vessels and dorsal longitudinal anastomotic vessels in zebrafish embryos following anisomycin treatment (n=30). Magnification, ×40. **P<0.01 vs. non-treated. HuOCSCs, human ovarian cancer stem cells; GFP, green fluorescent protein.