FIGURE 6.
Immunofluorescence microscopy of matrilin-4-transfected 293EBNA cells and mouse nasal cartilage sections. Total matrilin-4 and the cleavage site neo-epitope were detected with affinity purified antibodies raised against full-length matrilin-4 (anti-matn4) or the N-terminal neo-epitope of the cleaved C-terminal coiled-coil fragment of matrilin-4 (anti-matn4pc), followed by an Alexa 488TM-conjugated secondary antibody. To demonstrate the specificity, the anti-matn4pC antibody was preincubated with the same peptide that was used for immunization (anti-matn4pC inhibited). A–C, cells were transfected with StrepII-tagged matrilin-4 cDNA and grown on chamber slides. D–F, immunohistochemistry was performed on frozen tissue of newborn mice. Total matrilin-4 was detected throughout the nasal septum cartilage (ns). The neo-epitope antibody detects matrilin-4 in the perichondrium and weaker in cartilage. Arrows indicate nonspecific mucosa (mu) staining that is not affected by preincubation with the neo-epitope peptide used for antibody generation. The bars are 50 μm.