QUICK SEARCH:

[advanced]

	Author:		Keyword(s):

Home Help Feedback Subscriptions Archive Search Table of Contents

	Help viewing high resolution images
	Return to article

(Downloading may take up to 30 seconds.
If the slide opens in your browser, select File -> Save As to save it.)
Click on image to view larger version.

Fig. 3. Endogenous Abl kinases phosphorylate SHP-2 on Y580, and are required for sustained ERK activation in response to PDGF. (A) Serum-starved 10T1/2-EGFR cells were pretreated with STI571 (10 µM), stimulated with EGF, and lysates were probed with phospho-specific SHP-2 antibodies (Y580, Y542). Percentage phosphorylation is relative to total SHP-2 protein levels obtained from reprobed blots. (B) Percentage phosphorylation in lysates probed with phospho-SHP-2-Y580 antibody (as in A). Values are mean ± s.e.m. of three independent experiments. *P $≤$ 0.05 using a ratio paired t-test. (C) 10T1/2-EGFR or NIH3T3 cells, transfected with Abl or Arg siRNAs (40 nM), were serum-starved, stimulated with EGF (100 ng/ml; 10T1/2-EGFR) or PDGF-BB (12.5 ng/ml; NIH3T3) for 30 or 10 minutes, respectively, and lysates were probed with the indicated antibodies. SHP-2-Y580 phosphorylation in Abl or Arg siRNA-transfected cells was expressed as a percentage of SHP-2-Y580 phosphorylation observed in scrambled control-transfected cells and is relative to total SHP-2 protein levels obtained from reprobed blots. (D) Results from three independent experiments as in C. Values are mean ± s.e.m.*P $≤$ 0.05, ***P $≤$ 0.005 using a one-way ANOVA followed by a Bonferroni post-hoc test. (E) NIH3T3 cells, transfected with siRNAs, were serum-starved, stimulated with PDGF-BB (12.5 ng/ml) for 30 minutes, and phosphorylation of ERK1/2 was assessed by western blotting with a phospho-specific antibody [T(P)EY(P)]. ERK phosphorylation shown is relative to total ERK protein levels obtained from reprobed blots, and is expressed as a percentage of phosphorylation observed in scrambled control-transfected cells. (F) Results from three independent experiments as in E. Values are mean ± s.e.m.*P $≤$ 0.05, **P $≤$ 0.005, ***P $≤$ 0.001 using a one-way ANOVA followed by a Bonferroni post-hoc test.

Return to article