Figure 7.

Determination of the FN mutation responsible for the dominant-negative phenotype of FN matrix formation. (A) PCR products (7,114 bp) of full-length FN were obtained after amplification using a pair of primers and the first strand cDNA generated from F27mel tumor RNA. Cloning of both wild-type and mutant FN in a pcDNA3 expression vector. Wild-type and mutant FN were verified by DNA sequencing. (B) Immunostaining of FN matrices in 1143mel and derivative cell lines expressing wild-type FN, mutant FN, or empty vector. DAPI staining was used as a control for cell density.