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. Author manuscript; available in PMC: 2022 Jun 16.
Published in final edited form as: Neuron. 2021 May 24;109(12):1963–1978.e5. doi: 10.1016/j.neuron.2021.04.021

Figure 6. Promotion of Rapsn LLPS by Musk signaling.

Figure 6.

(A-E) Promotion of Rapsn LLPS by tyrosine phosphorylation. Rapsn-EGFP was treated with PP, CK2, or Src. (A) Phosphorylation levels of Rapsn-EGFP. (B, C) PP reduces and Src increases droplet formation. (B) Representative blots. (C) Quantitative data. ***, P < 0.001; One-way ANOVA. (D, E) PP reduces and Src increases Rapsn in pellets. (D) Representative silver staining. (E) Quantitative data. n = 3; ***, P < 0.001; One-way ANOVA.

(F-J) Enhanced Rapsn LLPS by Musk-induced tyrosine phosphorylation. Rapsn-EGFP was purified from HEK293T cells cotransfected with Musk or Musk-KD.

(F) Tyrosine phosphorylation of Rapsn. (G, H) Increased droplet formation by Musk cotransfection. ***, P < 0.001; One-way ANOVA.

(I, J) Increased Rapsn in pellets by Musk cotransfection. n = 3; ***, P < 0.001; One-way ANOVA.

(K-O) Reduced Rapsn LLPS by Y86F mutation. Rapsn or Y86F-EGFP was transfected into HEK293T cells alone or with Musk and purified for LLPS. (K) Tyrosine phosphorylation of Rapsn. (L) Representative images of droplets. (M) Quantification of droplet size in (L). n = 3; ***, P < 0.001; One-way ANOVA. (N) Reduced Y86F Rapsn in pellets. (O) Quantitative data in (M). n = 3; ***, P < 0.001; One-way ANOVA.

Data was shown as mean ± SEM; n = or > 3.