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. 2021 May 4;10:e66165. doi: 10.7554/eLife.66165

Figure 6. Mutant SHANK3 variants readily interact with cortactin, homer, and SAPAP1.

Figure 6.

(A) A representative western blot from co-IP assays of SHANK3 and cortactin from HEK293 cells is shown (detection SHANK3: rat-α-mRFP, 1 s; detection cortactin: mouse-α-GFP, 1 s; n = 4 independent experiments). No difference in binding of cortactin to ASD-associated SHANK3 mutants can be detected compared to the WT (one-way ANOVA with Bonferroni multiple comparisons test: p(WT/R12C)=1, p(WT/L68P)=1, and p(R12C/L68P)=1 at p=95%). (B) Representative western blot from co-IPs of SHANK3 and homer (detection SHANK3: rat-α-mRFP, 1 s; detection homer: mouse-α-GFP, 1.5 s; n = 4 independent experiments). No difference in binding of homer to the tested SHANK3 mutants can be found compared to the WT (one-way ANOVA with Bonferroni multiple comparisons test: p(WT/R12C)=1, p(WT/L68P)=1, and p(R12C/L68P)=0.99 at p=95%). (C) Representative western blot from co-IPs of SHANK3 and SAPAP1 (detection SHANK3: rat-α-mRFP, 9 s; detection SAPAP1: mouse-α-GFP, 2 s; n = 3 independent experiments). No difference in binding of SAPAP1 to the SHANK3 mutants can be detected compared to the WT (one-way ANOVA with Bonferroni multiple comparisons test: p(WT/R12C)=1, p(WT/L68P)=1, and p(R12C/L68P)=0.41 at p=95%). co-IP = co-immunoprecipitation, WT = wild type, ASD = autism spectrum disorders.

Figure 6—source data 1. Quantification of western blot data.