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Figure 5 | BMC Neuroscience

Figure 5

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

Figure 5

Upon knockdown of SHP-2 in C2C12 myotubes, AChR clusters are less stable. (A) pSUPER-, and pSUPER.shSHP2-transfected C2C12 myotubes were either left untreated (No Agrin), treated with agrin for 16 h (Agrin), or treated with agrin (16 h) followed by withdrawal of agrin for 8 h (Agrin+Withdr.). AChRs were identified by rhodamine-α-BTX staining and cells were analyzed by fluorescence microscopy. Scale bar, 40 μm. Transfected myotubes expressing EGFP indicated that transfection efficiency at the myotube level approached 100%. The data show that agrin induces AChR clustering both in pSUPER- and pSUPER.shSHP2-transfected myotubes. After the withdrawal phase, less clusters remain in the case of SHP-2 knockdown when compared to the control. (B) The stability of clusters was quantitated by setting the number of AChR clusters in agrin-treated myotubes (Agrin) to 100%, both for pSUPER-, and pSUPER.shSHP2-transfected myotubes. Clusters after the withdrawal period (Agrin + Withdr.) were then calculated accordingly. The absence of SHP-2 renders AChR clusters significantly less stable when compared to control transfected myotubes (means ± SEM, N = 30 from four similar experiments; *p < 0.05; two-tailed unpaired t test).

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