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Fig. 6 | Breast Cancer Research

Fig. 6

From: Nitric oxide deficiency and endothelial–mesenchymal transition of pulmonary endothelium in the progression of 4T1 metastatic breast cancer in mice

Fig. 6

Slit2–ROBO4–ROBO1-dependent signaling in orthotopic murine 4T1 breast cancer model. Slit2 (axon guidance molecule), ROBO4 and ROBO1 (roundabout 1 and 2) expression determined by western blot analysis in pooled samples (n = 6) corresponding to healthy control and 4T1 breast cancer-bearing mice from 1st to 5th week after 4T1 cancer cell inoculation (see Methods). Full-length Slit2 ~ 200 kDa (U) is proteolytically cleaved, giving rise to an N-terminal fragment ~ 140 kDa (N) and a C-terminal fragment ~ 55 kDa (C). Only full-length and N-terminal peptides can bind ROBO proteins. Therefore, representative picture of entire membrane probed with primary anti-Slit2 antibody presented; we are unsure about identity of the ~ 75 kDa band detected with anti-Slit2 antibody. Results presented as fold change vs control sample corresponding to healthy mice. Total protein after transfer was used as loading control. ROBO roundabout family of receptors, Slit axon guidance molecule

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