NES-ΔN-shootin1 disturbs the linkage between shootin1 and actin filament retrograde flow. (A) Schematic representation of the wild-type (WT) and ΔN- and ΔC-shootin1, and their abilities to interact with actin filament flow and inhibit the interaction between EGFP-shootin1 and actin filament flow. Numbers indicate amino acid numbers. (B and C) Fluorescent speckle images of EGFP-ΔN-shootin1 (B, left), and EGFP-shootin1 and mCherry-actin coexpressed in the same XTC fibroblast (C, left), with a time series of the indicated area of the cells at 10-s intervals (right). Myc-NES-ΔN-shootin1 was also overexpressed in the cell in C under the β-actin promoter. See Videos 8 and 9 . (D and E) Fluorescent speckle images of EGFP-shootin1 (left) in XTC fibroblasts and a time series of the indicated area of the cells at 10-s intervals (right). Myc-shootin1 (D) and myc-ΔC-shootin1 (E) were also overexpressed in the cells under the β-actin promoter. Arrowheads indicate speckles of EGFP-ΔN-shootin1 (B), mCherry-actin (C), and EGFP-shootin1 (D and E) moving retrogradely. Bar, 5 μm.