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. 2010 May 3;189(3):541–556. doi: 10.1083/jcb.200909113

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© 2010 Schoumacher et al.

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Figure 1. — Stages of BM breakage by invasive cancer cells. (A) Mesothelial BM. (left panels) x–y images showing the top of the BM immunostained for laminin (left) or collagen IV (right). (right panels) x–z sections of the corresponding x–y projection revealing the two BM layers present in the rat peritoneum. (B and C) Cells cultured atop of peritoneal BM stained with phalloidin–Alexa Fluor 488 (green) and DAPI (blue). BM was detected by laminin staining (red). Merged images are shown in the right column. HT29 (B) and HCT116 (C) cells are shown. (top) Stage 1, early stage of invasion characterized by degradation of the BM and formation of short invasive protrusions (invadopodia). (middle) Stage 2, intermediate stage of invasion and formation of long invasive protrusions (mature invadopodia). (bottom) Stage 3, late stage of invasion and infiltration of the cell on the other side of the membrane. Asterisks indicate sites of degradation and localization of invasive protrusions. (D) Time requirement for BM penetration by HCT116 cells. Black bars show the percentage of cells in stage 1. Light gray bars show the percentage of cells in stage 2. Dark gray bars show the percentage of cells in stage 3. Bars: (A and C) 5 µm; (B) 10 µm.