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. 2006 Aug 24;25(17):4050–4060. doi: 10.1038/sj.emboj.7601288

Figure 4.

Figure 4

Cholesterol does not induce AChR clustering and phosphorylation of MuSK and AChR β subunits. (A) Cholesterol or 1 nM agrin were added overnight to C2C12 myotubes. Cells were stained with rhodamine-α-BT and AChR clusters quantitated as in Figure 3. (B, C) C2C12 myotubes were treated with different doses of cholesterol, or with 1 nM agrin for 40 min, as indicated; C, untreated control. From cell lysates, MuSK was immunoprecipitated (B) or AChRs were precipitated using biotin-α-BT and streptavidin-agarose (Tox-P; C). Phosphotyrosine immunoblotting detected phosphorylation of MuSK and AChR β subunits. The identity of these phosphoproteins was confirmed by reprobing with MuSK- or AChR β-specific antibodies (not shown).