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. 2008 Feb 13;28(7):1688–1696. doi: 10.1523/JNEUROSCI.4130-07.2008

Figure 8.

Figure 8.

Suppression of NSF expression and mutation of NSF attenuate the agrin/MuSK signaling. A, Inhibition of agrin-induced MuSK phosphorylation by NSF shRNA and E329Q. C2C12 cells were transfected with NSF shRNA-491 or E329Q-NSF before stimulation with agrin (5 nm; 30 min). MuSK was immunoprecipitated (IP) and examined for tyrosine phosphorylation by immunoblotting with anti-phosphotyrosine antibody (top panel). The bottom panel shows equal amount of precipitated MuSK. B, Effect on Abl tyrosine phosphorylation. C2C12 cells were transfected with NSF shRNA-491 or E329Q before stimulation with agrin (5 nm; 30 min). Phosphorylation of Abl was examined with anti-Abl phosphorylation-specific antibody. C, Inhibition of Rac activation. Active Rac was purified by GST-PBD (Cdc42/Rac-binding domain of PAK fused to GST) immobilized on beads and revealed by Western blotting with anti-Rac antibody (top panel). The bottom panel shows equal amounts of Rac.