Journal: Oncogene

Article Title: Transformation and Stat activation by derivatives of FGFR1, FGFR3, and FGFR4.

doi: 10.1038/sj.onc.1203650

Figure Lengend Snippet: Figure 6 Activation of Stat1 and Stat3 by FGFR derivatives. (a) Lysates of 293T cells transfected as indicated were analysed by Western blotting using Phospho-Stat1 (Y701) antisera (top) or Stat1 antisera (bottom). (b) The same lysates in (a) were re- examined for phosphorylated Stat3 by Western blotting with Phospho-Stat3 (Y705) (top) or Stat3 antisera (bottom). Arrows in (a) and (b) indicate the positions of phosphorylated Stat1/3 and endogenous Stat1/3 in the lysates. Receptor expression levels were equivalent in each sample. (c) Cells were co-transfected with 46StRE-luciferase, a Stat-responsive reporter construct, and various FGFR derivatives, and subjected to luciferase assay. The graph shows the average relative luciferase units (RLU) for duplicate readings of samples from four independent experiments

Article Snippet: Primary antibodies were obtained from the following sources: Flg (C-15; FGFR1), FGFR3 (C-15), FGFR4 (C-16), SHPTP2 (C-18; Shp2), Stat1 (C-136) ± Santa Cruz Biotechnology; Phospho-MAPK (T202/Y204), Phospho-Stat1 (Y701), Phospho-Stat3 (Y705) ± New England Biolabs, Inc.; Mouse Anti-MAPK (ERK1+ERK2) ± Zymed; 4G10 (anti-phosphotyrosine) ± Upstate Biotechnology, Inc.; Stat3 ± Transduction Labs; PLC-g ± a kind gift of Jill Meisenhelder (Salk Institute); Secondary antibodies were obtained from the following sources: HRP-conjugated donkey anti-rabbit IgG and sheep anti-mouse IgG ± Amersham; HRP-conjugated Oncogene anti-goat IgG ± Santa Cruz Biotechnology and Sigma; Texas Red-conjugated goat anti-rabbit IgG ± Cappel/ICN; ̄uorescein-conjugated phalloidin ± Sigma.

Techniques: Activation Assay, Transfection, Western Blot, Expressing, Luciferase, Construct