UMSCC1 transductant cells were lysed with cold IP Lysis/Wash Buffer containing phosphatase inhibitor cocktails (set 1: Cat.
Total levels of Tks5, cortactin, and fascin proteins in the UMSCC1 transductant cells were assessed by Western blot analyses.
(54) Immunoprecipitation experiments of total tyrosine phosphorylated proteins in the UMSCC1 transductant cells were conducted to evaluate the overall tyrosine phosphorylation state of cortactin.
(54) For this assessment, UMSCC1 transductant cells were plated on the Alexa Fluor 405 gelatin-coated MatTek dishes, cultured for 4 hours, fixed, and stained by using total anti-cortactin and anti-phosphocortactin (pY421) antibodies.
(19,23,59-64) As we reported previously, the proliferation rates of stable transductant lines were not impaired by increased miR-375 expression, compared to the empty vector control lines.
Representative images of fluorescent matrix (A, D), tyrosine kinase substrate with 5 SH3 domains (Tks5) (B, E), and cortactin (C, F) immunostaining for UMSCC1 transductant cells.
Representative Western blots of total Tks5 (A), cortactin (B), and fascin (C) protein levels in UMSCC1 transductant lines.
Representative images of fluorescent matrix (A, D), cortactin (B, E), and phosphocortactin (pY421) (C, F) immunostaining for UMSCC1 transductant cells.
A, Representative images of protease arrays incubated with cell culture supernatants from UMSCC1 transductant cells.