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

FIGURE 3.

FIGURE 3.

Dynactin-4 requires ankyrin-B-binding activity for costamere localization of microtubules, dystrophin, and dystroglycan. A, N331A mutation of dynactin-4 impairs interaction with ankyrin-B (ankB) in yeast two-hybrid assays. B, shown are immunoblots (anti-HA) of HA-dynactin-4 and dynactin-4-N331A in transfected mouse TA muscles (as described under “Experimental Procedures”). C, single TA muscle fibers were transfected in vivo with a dynactin-4 siRNA and rescued with either wild type dynactin-4 or with dynactin-4-N331A (as described under “Experimental Procedures”). Rows from top to bottom, dynactin-4-siRNA-transfected muscle fibers (dynactin-4 detection with anti dynactin-4 antibody): middle row, dynactin-4 siRNA co-transfected with HA-wild type dynactin-4; bottom row, dynactin-4 siRNA cotransfected with HA-dynactin-4-N331A (middle and bottom rows, dynactin-4 detection with an anti-HA antibody). D, immunostaining for Arp1 in control, dynactin-4-, ankyrin-B-, and β2 spectrin-depleted muscle fibers (see “Experimental Procedures”). Scale bar, 5 microns.