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. 2019 Dec 9;219(1):e201902088. doi: 10.1083/jcb.201902088

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© 2019 Boyer et al.

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Figure 7. — TRIM67 colocalizes with TRIM9 and inhibits the TRIM9:VASP interaction. (A) Colocalization of GFP-TRIM9 and tagRFP-tagged TRIM67 in embryonic cortical neurons. (B) Quantification of colocalization between TRIM9 and TRIM67 (obs) compared with Fay-randomized controls (rand). Three experiments per treatment. n (cells) = 22 media, 11 netrin. (C) Coimmunoprecipitation of myc-tagged TRIM9 or TRIM67 from HEK293 cells showing coprecipitated GFP-VASP. (D) Quantification and individual data points of coprecipitated GFP-VASP relative to myc-tagged TRIM protein normalized to the VASP/TRIM67 ratio of each experiment. n (cultures) = 4 per genotype. Error bars denote SEM. (E) Coomassie-stained gel of recombinant GST or GST-EVH1 domain of VASP, used for pulldowns in F. (F) Pulldowns (PD) from embryonic mouse cortical lysate using either GST or GST-EVH1 domain of VASP, probed for endogenous TRIM67 and TRIM9. IB, immunoblot. (G) Quantification and individual data points of TRIM proteins precipitated by GST-EVH1 from lysates of indicated genotypes, normalized to WT levels. n (cultures) = 4 per condition. Error bars denote SEM. Data are presented as bar charts in D and G, as these datasets contain fewer than the five samples required to produce a box plot without interpolation. *, P < 0.05; ***, P < 0.005; n.s., P > 0.05. Box plots are minimum, Q1, Q2, Q3, maximum.