The human melanoma cell line A375P was infected with pBabe retroviruses encoding puromycin resistance or, in addition, cDNA for human integrin beta 6. Cells were selected in puromycin (1.25 ug/mL) followed by magnetic bead sorting, using 10D5 (mouse anti-avh6; Chemicon International), according to the manufacturer’s instructions (Dynal; Invitrogen). The αvβ6 expression can drift so recipients must check by flow cytometery after they have been in continuous passage, at least once a month. Use the magnetic beads above to re-select when this happens.
Useful model for studying the function and activity of beta 6 integrin when used as a matched pair with the parent A375 line. A375 cells endogenously express a variety of integrins, but not beta 6. These cell lines have been used to validate the activity and selectivity of AvB6-binding peptides. Matched pair of A375P melanoma cell lines stably transfected either a puromycin resistance gene or puromycin resistance + human integrin beta 6
DiCara et al. 2007. J Biol Chem. 282(13):9657-65. PMID: 17244604.
Barts Cancer Institute
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