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Fig. 5
SHG imaging reveals a defect in collagen thickening in the caudal fin of mmp9 morphants at 2days post amputation. (A) Amputation of the larval caudal fin results in a thickening of type I/III collagen fibers by 2dpw (top). Fiber thickening does not appear to occur in the mmp9 morphants (MO1) by 2dpw (bottom). (B) Scoring system for fiber width, from 1 to 3 (top). A significant defect in collagen fiber thickness is seen in the mmp9 morphants at 2dpw (bottom). (C) Schematic illustrating the quantification of SHG images. All fibers were measured and the number of fibers above a given threshold width (i.e. 7 pixels) can be determined, see Materials and Methods. (D) Quantification of the number of fibers >7 pixels in diameter per caudal fin validates a defect in collagen fiber thickening in the mmp9 morphants at 2dpw. **P<0.01, ***P<0.001 and ****P<0.0001. Scoring in B from a single-blind analyzer from pooled experiments performed in triplicate. Data in D are from experiments performed in quadruplicate and scored by a single-blind analyzer. Scale bars: 50µm in A; 20µm in B. Experimental numbers for B and D: no wound Control MO=21, no wound mmp9 MO=25, wounded Control MO=31, wounded mmp9 MO=32.