Figure 8.

Plasma FN matrix assembly defect and its increased degradation in Tgm2^−/−;F13a1^−/− bmMSC cultures. (a). BMMSCs isolated from 2-month-old mice were grown with biotinylated plasma FN (bpFN) and assembly of DOC-soluble matrix was examined by Western blotting. A decrease is seen in DOC-soluble bpFN matrix in Tgm2^−/−;F13a1^−/− mouse cultures. A smaller pFN fragment is also seen in Tgm2^−/−;F13a1^−/− cultures indicative of increased degradation. Western blot image is representative of two separate cell culture experiments. (b) Double-null bmMSCs show no altered proliferation under regular media or adipogenic media as per MTT assay. Triplicate analysis from two separate experiments is presented. (c) Immunofluoresence analysis of bpFN assembly (green), cell density (DAPI, blue) proliferation (Ki67+DAPI)(pink) around bpFN. WT bmMSCs assemble bpFN into clear rounded patches that are associated with cell nodules. Tgm2^−/−;F13a1^−/− bpFN matrix lacks this rounded shape and nodular configuration of the cells. Cell proliferation appears normal. Immunofluoresence images are representative of two separate cell culture experiments. NS, not significant. Scale bars, 10 μm