Additional file 7: Figure S5.

Lung metastasis assay after modulation of miR30 expression. 4T1 mammary cancer cells were transiently transfected with miR-30a family KD, Pre-miR-30a, and miR-159 KD control oligos. Next, cells were injected in the mammary fat pad of BALB/c mice and tumor growth was followed during 21 days (experiment shown in Figure 5C). Lungs obtained after the different conditions were minced, dissociated, and cells were suspended in medium containing 6-thioguanine and plated in culture dishes. After 14 days, cells were fixed by methanol and stained with methylene blue solution (A). All blue colonies were counted, one colony representing one clonogenic metastatic cell (B).

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Ouzounova et al. BMC Genomics 2013 14:139   doi:10.1186/1471-2164-14-139