Figure 8: CRN7 regulates cellular processes via N-WASP.
From: Novel Coronin7 interactions with Cdc42 and N-WASP regulate actin organization and Golgi morphology
(a) Domain structure and deletion fragments of N-WASP. The positions of the amino acids are indicated. (b) Co-IP using GFP microbeads to precipitate GFP-N-WASP and deletion fragments ΔWA, WH1 and WH1-GBD from HEK293T cell extracts followed by immunoblot analysis using mAb K37-142-1 to detect CRN7. GFP mAb antibody K3-184-2 detected immunoprecipitated N-WASP polypeptides. (c) Evaluation of the binding shown in (b) represented as a ratio of co-IP (CRN7) to IP (N-WASP fragments) (n = 4 independent experiments, mean ± SD; *P < 0.05). (d) F-actin content determined from phalloidin-stained cells. Mean F-actin intensity values derived from z-stack images (n = 20 cells each, 2 independent experiments; *P < 0.05). AU, arbitrary units. Data shown as mean ± SD. (e) Quantification of cell spreading area at 30 and 60 min. Cells stained with phalloidin were evaluated (n = 15 cells each, each time point, 2 independent experiments; n.s., not significant). (f) Percentages of cells with fragmented or compact Golgi represented as stacked columns (n = 28 cells each, 2 independent experiments).
