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. 2012 Jun 15;7(6):e39065. doi: 10.1371/journal.pone.0039065

Figure 6. WFA has no effect on the in vitro assembly of human recombinant vimentin.

Figure 6

(A) Recombinant human vimentin (0.2 mg/ml) was assembled for 10 min at 37°C in (i) 50 mM NaCl; (ii) with 0.25% DMSO; (iii) with 50 μM WFA; and (iv) for 30 min with 50 μM WFA at a protein concentration of 0.5 mg/ml. The filaments were fixed with glutaraldehyde and visualized by negative stain electron microscopy. The arrows in (ii) indicate lateral annealing and apparent fusion of individual filaments. (scale bars, 0.2 μm). (B) Viscometric analysis of vimentin assembly in the absence (ctrl) and presence of 50 μM WFA at 37°C in 50 mM NaCl. (C) Centrifugation assay of vimentin assembled in the absence (c) and presence of WFA (w). VIF were assembled for the indicated times (5 to 15 min) in 160 mM NaCl then centrifuged for 5 min at 10 psi in an Airfuge. Samples were separated by SDS-PAGE and stained with Coomassie. The position of vimentin is indicated (55 kDa).