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Fig. 3. Anillin and RacGAP50C interact in vitro. (A) At the left is shown a schematic representation of the full-length Anillin protein and of the several fragments used for in vitro binding assays. The PH domain and myosin (My)- and actin (Ac)-binding regions are indicated. The thick grey line marks the region showing the highest homology among different species (Anillin Homology Domain, AHD). The autoradiograph of the pull-down assay along with a Coomassie-stained gel (CBB) showing the amounts of bait proteins used for the binding assay are shown at the right. The top bands in the Coomassie gel correspond to the full-length GST-tagged baits. (B) Subcellular localization of GFP-tagged Anillin fragments. A schematic description of the fragment tagged with GFP is shown at the left and their relative subcellular localization at the right. The cells were fixed and stained to detect tubulin (red in merged panels), GFP (green in merged panels) and DNA (blue in merged panels). The arrowhead marks the spindle midzone localization of Ani_410-1104. (C) At the left is shown a schematic representation of the full-length RacGAP50C protein and of the several fragments used for in vitro binding assays. The GAP and C1 domains, and Pav-KLP (Pav)- and Pebble (Pbl)-binding regions are indicated. The autoradiographs of the pull-down assays are shown at the right. The amount of GST and GST-A4 proteins used is the same as in A. Scale bar: 10 µm.

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