Supplemental Figure 1
-
Fig. S1. Examination
of Other MMPs in Mmp13–/– long bones. In situ
hybridization for Mmp14, Mmp8 and Mmp2
mRNA in 1-week-old metatarsals of wild-type and Mmp13–/– mice, showing no alteration in
expression of Mmp14, Mmp8 and Mmp2 in Mmp13–/–
endochondral long bones. Immunostaining for MMP9 in 1-week-old metatarsals of
wild-type and Mmp13–/–
mice showing an expanded zone of MMP9 protein localization in the matrix
surrounding the most terminal hypertrophic chondrocytes (HC) in Mmp13–/– mice. This is caused by the
persistence of the protein with the slowed degradation of the ECM. In situ
hybridization analyses for Mmp9 did
not reveal a difference in expression levels in mutants compared with wild type
(data not shown). For MMP9 immunohistochemistry, tissues were fixed briefly in
4% paraformaldehyde, decalcified in EDTA, frozen in OCT, sectioned at 5 mm using a cryostat and briefly post-fixed in
4% paraformaldehyde. Slides were deglycosylated in Chondroitinase ABC and
rabbit anti-MMP9 antibody was used as described previously (Vu et al., 1998).
Scale bars: 100 mm
