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Fig. 6. The LIM2 and LIM3 domains are crucial for Hic-5 to be localized to stress fibers. (A) The HA-tagged Hic-5 mutants in which each of the LIMs was disrupted by point mutations (mL1-mL3) or a deletion (delL4). The expression plasmids for each mutant have been previously described (Shibanuma et al., 2003). (B) The expression plasmids were transfected into mouse embryo fibroblast cells; 24 hours later, western blotting was carried out using the antibody against the HA tag. (C) Mouse embryo fibroblast cells were transfected with each expression vector, subjected to cyclic stretching (+) or not (–) and immunostained with the 12CA5 antibody against the HA tag. (C, left) Staining of the Hic-5 mutants (red) with the 12CA5 antibody to the HA tag; (C, middle) F-actin stained by FITC-conjugated phalloidin (green); (C, right) merged images. The arrow indicates the direction of stretch. Bars, 5 µm.

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