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Figure 3 | BMC Neuroscience

Figure 3

From: Tyrosine phosphatases such as SHP-2 act in a balance with Src-family kinases in stabilization of postsynaptic clusters of acetylcholine receptors

Figure 3

Protein tyrosine phosphatase SHP-2 increasingly associates with MuSK upon agrin stimulation. (A) C2C12 myotubes were treated with 0.5 nM agrin for 40 min. Following MuSK immunoprecipitation, immunoblotting for SHP-2 and MuSK was carried out, followed by reprobing for phosphorylation of MuSK. Levels of SHP-2 co-precipitated with MuSK were quantitated (normalized for the amount of precipitated MuSK) as percentage of No Agrin control. Co-immunoprecipitation of SHP-2 with MuSK indicates its specific association with MuSK, which increases significantly upon agrin-induced MuSK phosphorylation (means ± SEM, from seven experiments; * p < 0.05; two-tailed unpaired t test). As a control, the MuSK antibody was omitted in the immunoprecipitation, leading to no substantial signal in the Western blots (not shown). (B)COS cells were co-transfected with SHP-2-Flag and MuSK-myc constructs. Cells were then fixed and stained with anti-Flag/Alexa488 and anti-myc/Alexa546 antibodies. SHP-2 localizes throughout the whole of the cytoplasm, while MuSK localization is concentrated along the plasma membrane. White arrowheads in the overlay indicate a colocalization of MuSK and SHP-2 along membrane ruffles on the plasma membrane. Scale bar, 40 μm.

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