Figure 6. CRP2 is required for ECM degradation.

A. Gelatin degradation assay. Control sh- and shCRP2a and shCRP2b cells were plated on Cy3-conjugated gelatin for 18 h, fixed and stained with DAPI and Acti-stain^TM 670 phalloidin (left panels). Gelatin degraded areas appear as dark punctuate in the fluorescent background (right panels). B. Same as A for an shCRP2a-derived cell line in which CRP2 expression was restored by expressing an shRNA-resistant CRP2 coding sequence (shCRP2a/rescue), and related controls, i.e. sh- and shCRP2a cells lines transduced with an empty vector (sh-/empty and shCRP2a/empty, respectively; see also Supplementary Figure 4). C. and D. Quantitative analyses corresponding to the experiments shown in (A) and (B) Actively ECM degrading cells were scored and expressed as percentage of the total cell population (C). A degradation index corresponding to the ratio of degraded gelatin surface per cell was calculated (D). The data originate from at least three independent experiments (n ≥70 cells). Error bars denote standard error. Significant levels: *: p < 0.0001. Bars = 20 μm.