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Figure 1 | Journal of Molecular Signaling

Figure 1

From: Direct RNA sequencing mediated identification of mRNA localized in protrusions of human MDA-MB-231 metastatic breast cancer cells

Figure 1

Trans- migration of MDA- MB- 231 protrusions in a modified Boyden chamber assay. (A) Immunofluorescence image showing the typical morphology of human breast cancer MDA-MB-231 cells. The cellular cytoplasm was stained for α-Tubulin (α-Tub, green) and nuclei by DAPI (Blue). Scale bar 20 μm. (B) Schematic drawing showing the principle of the Boyden chamber set-up for isolation of migrating protrusions. Cells were grown on a 1 μm pore size micro porous membrane allowing trans-migration of only the cell protrusions. The lower side of the membrane is coated with ECM protein. (C) Protrusions of MDA-MB-231 cells migrated through a 1 μm micro porous membrane after 24 h assay time. α-Tubulin (green) and DAPI staining (blue) is shown without and after wipe (wipe) to remove cellular material present on the upper side of membranes. Examples of protrusions present on the Boyden chamber membrane lower side are indicated by arrowheads. Scale bar 100 μm. (D) Histone H3 is absent from the Boyden chamber lower side protrusion fraction. Western blotting was performed with histone H3 antibody and α-Tubulin antibody on cell body fraction (CF) and protrusion fraction (PF) protein samples originating from pooled material from three independent Boyden chambers.

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