Journal: Biochimica et biophysica acta

Article Title: Persistence length of fascin-cross-linked actin filament bundles in solution and the in vitro motility assay.

doi: 10.1016/j.bbagen.2014.01.012

Figure Lengend Snippet: Fig. 1. Structural characteristics of fascin-mediated actin bundle. (A) Schematic diagram of actin bundles showing the bundling of actin filaments (F-actin; red) mediated by fascin (blue). Molecular weights of fascin and actin in monomer form are 57 kDa and 42 kDa, respectively. (B) Actin-bundling assay of rhodamine-phalloidin-stabilized fascin–actin bundles using low-speed centrifugation. Pictures of SDS-PAGE gels visualized by Coomassie staining for the final fascin:actin molar ratios (1:7, 1:4 and 1:2) for pellets (pellet) and supernatant (supernatant) after low-speed centrifugation and for F-actin alone (30 pmol) and fascin alone (20, 40 and 60 pmol) before centrifugation (control).

Article Snippet: Histidine (His)-tagged fascin was purchased from Novus Biologicals (NBP145313, Novus Europe, UK), with a concentration of 0.89 ± 0.29 mg/ml (mean ± 95% confidence interval) according to the Bradford assay.

Techniques: Centrifugation, SDS Page, Staining, Control

Journal: Biochimica et biophysica acta

Article Title: Persistence length of fascin-cross-linked actin filament bundles in solution and the in vitro motility assay.

doi: 10.1016/j.bbagen.2014.01.012

Figure Lengend Snippet: Fig. 2. Fluorescent images of actin filaments (F-actin) (A) and fascin–actin bundles (B; fascin:actin ratio = 1:2) in solution. While F-actin is flexible and curved, fascin– actin bundles are nearly straight and with low flexibility. Scale bars: 10 μm. Details of the image processing are given in the Supplemental Information.

Article Snippet: Histidine (His)-tagged fascin was purchased from Novus Biologicals (NBP145313, Novus Europe, UK), with a concentration of 0.89 ± 0.29 mg/ml (mean ± 95% confidence interval) according to the Bradford assay.

Techniques:

Journal: Biochimica et biophysica acta

Article Title: Persistence length of fascin-cross-linked actin filament bundles in solution and the in vitro motility assay.

doi: 10.1016/j.bbagen.2014.01.012

Figure Lengend Snippet: Fig. 3. Image sequences depicting movement of actin filaments (F-actin) (A) and fascin:actin bundles (fascin:actin molar ratio = 1:2) (B) when they are sliding on heavy meromyosin (HMM) for 15 s in the in vitro motility assay. Images represent snapshots (0.5 s exposure time) obtained at 5 s interval. Different arrows denote the tip of individual F-actin and actin bundles during their sliding on HMM. Scale bars: 10 μm. Details of the image processing are given in the Supplemental Information.

Article Snippet: Histidine (His)-tagged fascin was purchased from Novus Biologicals (NBP145313, Novus Europe, UK), with a concentration of 0.89 ± 0.29 mg/ml (mean ± 95% confidence interval) according to the Bradford assay.

Techniques: In Vitro, Motility Assay

Journal: Biochimica et biophysica acta

Article Title: Persistence length of fascin-cross-linked actin filament bundles in solution and the in vitro motility assay.

doi: 10.1016/j.bbagen.2014.01.012

Figure Lengend Snippet: Fig. 5. Cosine correlation functions plotted against contour length s along filaments or bundles in solution (A) and against path length s from trajectory of filaments or bundles propelled by heavy meromyosin (HMM) on the surface in the in vitro motility assay (B). Experimental data were plotted for actin filaments (F-actin) (open circle) and fascin–actin bundles in the fascin:actin molar ratios of 1:7 (closed circle), 1:4 (open square) and 1:2 (closed triangle). Exponential fits were obtained using non-linear regression.

Article Snippet: Histidine (His)-tagged fascin was purchased from Novus Biologicals (NBP145313, Novus Europe, UK), with a concentration of 0.89 ± 0.29 mg/ml (mean ± 95% confidence interval) according to the Bradford assay.

Techniques: In Vitro, Motility Assay

Journal: Biochimica et biophysica acta

Article Title: Persistence length of fascin-cross-linked actin filament bundles in solution and the in vitro motility assay.

doi: 10.1016/j.bbagen.2014.01.012

Figure Lengend Snippet: Fig. 6. Effect of fascin and heavy meromyosin (HMM) induced sliding on persistence lengths of actin filaments (F-actin) and fascin–actin bundles. Persistence lengths in the in vitro motility assay (LpM; closed squares) and in solution (Lp; open squares) versus the fascin:actin molar ratio. Inset: LpM versus Lp for the range of contour lengths and path lengths between s = 0 μm to s = 15 μm. The quantities Lp and LpM were obtained from the exponential fits using the cosine correlation function, as shown in Fig. 5A and B. The straight lines were fitted to the experimental data using linear regression analysis. Lp and LpM for actin bundles were considerably larger than for F-actin and increased in proportion to the increased fascin:actin ratios.

Article Snippet: Histidine (His)-tagged fascin was purchased from Novus Biologicals (NBP145313, Novus Europe, UK), with a concentration of 0.89 ± 0.29 mg/ml (mean ± 95% confidence interval) according to the Bradford assay.

Techniques: In Vitro, Motility Assay

Journal: Biochimica et biophysica acta

Article Title: Persistence length of fascin-cross-linked actin filament bundles in solution and the in vitro motility assay.

doi: 10.1016/j.bbagen.2014.01.012

Figure Lengend Snippet: Fig. 8. Image series showing HMM-driven movement of fascin–actin bundles, each having short, apparently individual, actin filament at the front end as indicated by arrows. Actin filaments at the front end of bundles are shorter (A) and longer (B) than the thick part of bundles. In (B) the position of the single filament at the leading end at time 0 s is illustrated by yellow full lines in the figures at times 3 s and 6 s. The position of single filament at front at time 3 s is illustrated by dashed yellow line in figure at time 6 s. Scale bar: 10 μm. Details of the image processing are given in the Supplemental Information.

Article Snippet: Histidine (His)-tagged fascin was purchased from Novus Biologicals (NBP145313, Novus Europe, UK), with a concentration of 0.89 ± 0.29 mg/ml (mean ± 95% confidence interval) according to the Bradford assay.

Techniques: