Fig. 8. Effects of Hic-5, paxillin and CRP2 on collagen-gel contraction. (A) The Tet-Off/LD1mhic-5 (top) and the MEF/Tet-Off/paxillin (bottom) cell lines incubated with (Dox +) or without (Dox –) doxycycline for 24 hours were embedded in collagen gels and the gel contraction was assessed by measuring the diameters at each indicated time. The means and standard deviations of four parallel measurements are shown. (B, top) The Tet-Off/LD1mhic-5 cell line incubated with (Dox +) or without (Dox –) doxycycline for 24 hours was infected with the retrovirus of CRP2 (CRP +) or control virus (CRP –) for the next 24 hours and then embedded in collagen gel. The gel contraction was assessed as in A and the pictures were taken 56 hours after floating the collagen gels (C). (B, bottom) Expression plasmids encoding the mL2 and mL3 Hic-5 mutants were introduced into SVS30 smooth-muscle cells by electroporation and, 24 hours later, the cells were embedded in collagen gels and observed as in A. (D) Intracellular distribution of Hic-5 and CRP2 in the cells embedded in collagen gel. The cells in collagen gel were prepared as in C and stained with anti-Hic-5 (red) and anti-Flag polyclonal (CRP2) (green) antibodies, and DAPI (blue). Bar, 10 µm.