FIGURE 4.

Effect of mutation in L1 (L21A) and/or L2 (L307A) on subcellular localization of MRTF-A. A, COS-7 cells were transfected with expression plasmids for the indicated FLAG-tagged MRTF-A under serum-stimulated conditions. For the final 20 h, the cells were cultured under serum-starved conditions. The cells were then stained with anti-DYKDDDDK (FLAG) antibody (red) and Hoechst 33258 (blue). Representative images from at least three independent experiments are shown. Scale bar = 20 μm. B, the images were quantified as described under “Experimental Procedures.” N, nuclear accumulation; NC, diffuse distribution in the nucleus and cytoplasm; C, cytoplasmic localization. Statistical differences were calculated using Student's t test. *, p < 0.05 versus wild-type MRTF-A.