Figure 7.

Knocked down MEGF11 in the mouse 4T1 mammary cancer cell line decreased tumour weight and circulating tumour cells. Using the spontaneously occurring mouse mammary tumour 4T1 cell line as an in vivo metastatic model, 4T1 cells (1×10⁷/0.1 mL PBS) of wild type MEGF11 (n = 9) and ΔMEGF11 4T1 (n = 8) were orthotopically injected into two fat pads (left upper and right lower mammary glands). After two weeks, the 4T1-bearing mice were anaesthetized, and blood cells were collected and centrifuged with Ficoll-Paque (density: 1.084). The tumour weight (a) was measured. The implanted tumours were homogenized and analysed by Western blot (b,c). The peripheral mononuclear cells were harvested for primary cultures, and circulating 4T1 cells were selected with 6-thioguanine (60 µM) (d), which was analysed by Fisher’s exact test, followed by quantification by agar assay (e,f). A p value <0.05 indicates statistical significance in ΔMEGF11 4T1 cells compared to the wild type by Mann-Whitney U test (*) or by Student’s t test (#). NC, negative control; PC, positive control.