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. 2012 Apr 2;7(4):e34677. doi: 10.1371/journal.pone.0034677

Figure 6. NESH interacts directly with filamentous actin via its N-terminal region, but not with monomeric actin.

Figure 6

(A) Schematic diagram showing representations of full-length NESH (amino acids 1–367), N-term (N-terminal half, amino acids 1–229) and C-term (C-terminal half, amino acid 221–367). (B) GST pull-down assays were performed to verify the interaction between NESH and monomer G-actin. GST-fused NESH proteins were incubated with purified monomeric G-actin and then pulled down with glutathione Sepharose beads, after which the bound proteins were detected with anti-actin antibody. GST-SPIN90-C-term served as a positive control. (C) F-actin co-sedimentation assays. Purified NESH proteins were incubated with polymerized F-actin. After separating the supernatant (S) and pellet (P) by ultracentrifugation, co-sedimented proteins were detected by Coomassie Brilliant Blue staining. (D) NESH N-term and C-term in F-actin co-sedimentation assays. Note that NESH N-term only interacts with F-actin.