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Fig. 3. T-Plastin and ABD1 increase in actin tails concomitant with a significant increase in actin content and partial displacement of cofilin. (A) Quantification of actin and T-plastin or ABD1 in actin comets. Beads were incubated in HeLa-cell extracts supplemented with 1 µM Alexa-568-labeled actin in the presence of 1.1 µM T-plastin (middle) or ABD1 (bottom), or their absence (control, top); actin is shown on the left. Samples were immunostained with anti-plastin primary antibody and Alexa-488-coupled secondary antibody (right). Graphs indicate the integrated fluorescence intensities of actin and T-plastin in the comets, measured at 2 µm from the bead. Each bar represents the mean±s.d. of 25 measurements. Asterisks (**) indicate results that differ significantly from the results obtained with the control (*, P<0.0031; **, P<0.0063). (B) Quantification of cofilin/actin ratio in actin comets. Beads were incubated in extracts and processed as described in A, with the exception that anti-cofilin antibody was used for staining (middle). Representative fluorescence images are shown. Graphs indicate the ratios of the integrated fluorescence intensities of cofilin/actin in the comets, measured at 2 µm distance from the bead.
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