Fig. 6

WSB-1 promotes metastatic potential in vivo MDA-MB-231 cells were stably transfected with WSB-1 (shRNA WSB-1) or non-targeting shRNA (shRNA NT) constructs. The WSB-1 shRNA construct #1 was selected, as it led to the most efficient WSB-1 knockdown (Supplementary Figure 10). a WSB-1 protein levels were assessed by Western blotting. b and c Histograms represent VEGFA mRNA expression and VEGF secretion as previously described, average of n = 3 experiments. d Mammospheres were established from shNT and shWSB-1 MDA-MB-231 cells. Histogram represents the average mammosphere diameter per condition per individual experiment. Representative images are shown (n = 4). e–g 5 × 10⁵ cells were injected into the tail vein of female CD-1 nude mice (n = 6 per group). After 15 weeks, mice were sacrificed and lung tissue was collected and fixed. For each animal, the right lung was sectioned into 10 µm sections and a total of 10 equally distanced sections were H&E stained, imaged (10× objective), and scored for presence of metastatic foci. The metastatic lesions were also measured. The total number of metastatic foci per lung per group is represented in the scatter plot (e), and the histogram represents the average area occupied by metastatic lesions in relation to total lung area (f). Representative example of a section of each group is shown in (g). Arrowheads indicate the location of metastatic lesions. *p < 0.05