Immunofluorescence localization of phosphotyrosine containing proteins in RSV-transformed mouse fibroblasts

Pier Carlo Marchisio, Maria Flavia Di Renzo, Paolo M. Comoglio

Research output: Contribution to journalArticlepeer-review


The localization of phosphotyrosine-containing proteins (P-TYR proteins) was studied by immunofluorescence microscopy employing affinity-purified azobenzyl phosphonate (ABP) antibodies, which specifically cross-react with phosphotyrosine. In Rous sarcoma virus (RSV)-transformed fibroblasts, after fixation followed by permeabilization with nonionic detergents, ABP antibodies gave a diffuse staining of the cytoplasm and specifically decorated restricted areas of the ventral plasma membrane corresponding to adhesion plaques, identified by interference reflection microscopy and staining with anti-vinculin serum. Specific decoration was also observed at the level of cell-cell contacts and at the tips of filopodial protrusions. Control non-transformed 3T3 mouse fibroblasts were not significantly stained by ABP antibodies. These findings show that, in RSV-transformed cells, proteins phosphorylated at tyrosine residues are found at cell-substratum and cell-cell contacts.

Original languageEnglish
Pages (from-to)112-124
Number of pages13
JournalExperimental Cell Research
Issue number1
Publication statusPublished - 1984

ASJC Scopus subject areas

  • Cell Biology


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