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Figure 2


Fig. 2. Interaction of the AF6i3 C-terminus (AF6i3 CT) with F-actin. (Aa) Schematic representation of the AF6i3 CT constructs used in F-actin co-sedimentation assay (GST-tagged) and immunofluorescence (HA-tagged). (Ab,c) F-actin co-sedimentation assay performed with the GST fusion protein of (b) the whole AF6i3 CT (aa 1588-1743) or (c) the two C-terminal fragments (aa 1588-1666 and 1667-1743). F-actin-containing pellet (P) and the supernatant (SN) were analyzed on a denaturing protein gel and stained with Coomassie Blue. GST-fusion proteins or GST alone (arrows) and actin (arrowhead) are annotated. (B) Colocalization of the HA-tagged AF6i3-CT proteins (anti-HA antibody, green) and F-actin (red) in HeLa cells is depicted in merged pictures (right column). Bar, 20 µm. (C) Colocalization of endogenous (AF6; anti-AF6 antibody) and overexpressed full-length AF6i3 protein (myc-AF6i3; anti-myc antibody) (green) with F-actin (red). Bar, 20 µm.