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. 2010 Dec 25;286(9):7370–7378. doi: 10.1074/jbc.M110.187831

FIGURE 1.

FIGURE 1.

Ankyrin-B requires dynactin-4 binding activity to promote association of microtubules, dystrophin, and dystroglycan with costameres. A, ankyrin-B domains (MBD, membrane binding domain; SB, spectrin binding domain; DD, death domain; ankB, ankyrin-B; C′-term, C′-terminal). The ankyrin-B-DD1320AA mutation (red) resides in an exposed loop in the UPA domain modeled from the UNC5b cytoplasmic domain structure (20). B, DD1320 is required for ankyrin-B-dynactin-4 but not β2 spectrin interaction in a yeast two-hybrid assay (as described under “Experimental Procedures”). C, representative immunoblot with HA antibody of homogenates of TA muscles depleted of endogenous ankyrin-B and transfected with either HA-ankyrin-B or HA-ankyrin-B-DD1320AA (as described under “Experimental Procedures”). D, single TA muscle fibers transfected in vivo with ankyrin-B siRNA and rescued with either wild type ankyrin-B or with ankyrin-B-DD1320AA (as described under “Experimental Procedures”). Rows (top to bottom), untransfected control, ankyrin-B siRNA (in both ankyrin-B staining is with ankyrin-B antibody), ankyrin-B siRNA co-transfected with HA-tagged ankyrin-B, and ankyrin-B siRNA co-transfected with HA-tagged ankyrin-B-DD1320AA (in both ankyrin-B staining is with an HA antibody). Scale bar, 5 microns.