src family kinase y416  (Cell Signaling Technology Inc)

Journal: Scientific Reports

Article Title: The Phosphorylation and Distribution of Cortactin Downstream of Integrin α9β1 Affects Cancer Cell Behaviour

doi: 10.1038/srep28529

Figure Lengend Snippet: ( A ) WB of G361 TCL from cells in suspension or after adhesion to A12-Dis, in the absence or presence of Mn 2+ (1 mM). ( B ) WB of RD and G361 TCL from cells in suspension or after adhesion to TNfn3RAA, in the absence or presence of Mn 2+ (1 mM). ( C) Quantitation of ( B ) for relative levels of cortactin Y470 and Y421 tyrosine phosphorylation after Mn 2+ treatment. Data from untreated NC cells were set at 1.0. Data were analysed using Mann-Whitney Rank Sum test. ( D ) WB of TCL from G361, A375 and RD cells after adhesion to A12-Dis in the absence or presence of Mn 2+ (1 mM). ( E ) G361 cells were transfected with NC, Arg, Yes, or Crk siRNA, and changes in cortactin tyrosine phosphorylation of Y421 and Y470 upon knockdown were evaluated by WB. ( F ) WB of TCL from SYF cells and SYF cells re-expressing Yes (SYF-Yes) that were either left untreated or subjected to Mn 2+ treatment. This WB also shows that the antibodies recognising phosphorylated cortactin Y421 and Y470 discriminate phosphorylated from non-phosphorylated cortactin. Blots were analysed and probed for pTyr (4G10), cortactin, pY421 cortactin, pY470 cortactin, pY416 SFK, v-Src, Arg, Yes, Crk, and actin, as indicated.

Article Snippet: The following rabbit polyclonal antibodies were used: anti-cortactin (ab11066 from Abcam, for G361 lysates) and anti-cortactin phospho-Y470 (ab51703, from Abcam, or sc-101661 from Santa Cruz), anti-cortactin phospho-Y421 and anti-Src family kinases (SFK) phospho-Y416 (both from Cell Signaling), anti-arg, anti-Pyk2, anti-FAK (all from Millipore), anti-FAK phospho Y397 (Invitrogen), and anti-fibronectin (R2/7, made against bovine plasma fibronectin ).

Techniques: Quantitation Assay, MANN-WHITNEY, Transfection, Expressing

Journal: Scientific Reports

Article Title: The Phosphorylation and Distribution of Cortactin Downstream of Integrin α9β1 Affects Cancer Cell Behaviour

doi: 10.1038/srep28529

Figure Lengend Snippet: Representative confocal micrographs of G361 cells seeded on A12-Dis. Cells were stained for cortactin (cort), pY421 cortactin (pY421), pY470 cortactin (pY470), pY416 SFK (pY416), α9β1 (red), and F-actin (blue). Arrowheads indicate focal adhesion structures. Scale bars = 10 μm.

Article Snippet: The following rabbit polyclonal antibodies were used: anti-cortactin (ab11066 from Abcam, for G361 lysates) and anti-cortactin phospho-Y470 (ab51703, from Abcam, or sc-101661 from Santa Cruz), anti-cortactin phospho-Y421 and anti-Src family kinases (SFK) phospho-Y416 (both from Cell Signaling), anti-arg, anti-Pyk2, anti-FAK (all from Millipore), anti-FAK phospho Y397 (Invitrogen), and anti-fibronectin (R2/7, made against bovine plasma fibronectin ).

Techniques: Staining