Fig. 3

Nalbulphine decreases breast cancer stem-like properties in vivo. a Representative tumor image (left) from Ctrl and Nal mice. Growth of Ctrl and Nal MDA-MB-231 tumors in mice (n = 5). b Body weight growth curves of mice over 30 days following tumor implantation (n = 5). c, d ALT, AST (c), CRE and BUN (d) levels in serum of Ctrl or Nal mice (n = 5). e Representative spheroid images derived from primary Ctrl and Nal tumor cells (left) (n = 5); Scale bar, 50 μm. Mammosphere sizes (upper right) and numbers of mammospheres (bottom right) (d > 50 μm) were shown. f Primary cells were extracted from excised Ctrl and Nal tumors. Levels of indicated proteins were determined by western blot (n = 5). g Representative IHC staining for indicated proteins in Ctrl and Nal MDA-MB-231 tumor tissue (n = 5); Scale bar, 50 μm. h Serial diluted Ctrl MDA-MB-231 tumor cells were subcutaneously inoculated at four different sites in each mouse. Statistical analysis of tumorigenicity with indicated cell numbers and treatments is shown (n = 5). Data are presented as mean ± SEM. p-value was determined by Student’s t-test and ANOVA (a and e) (*p < 0.05, **p < 0.01, ***p < 0.001). See also Additional file 4: Figure S3