FIGURE 2.

MLCK-deficient SMCs display enhanced migration. Explants of jejunal smooth muscle tissues were cultured on gelatin-coated dishes, and the outgrowth cells were photographed at different time points. A, typical morphologies of control and MLCK-deficient SMCs cultured for 6 days are presented. Positive explants are defined as the explants with ≥1 cell outside after culture. The percentages of the positive explants of control (n = 24–38) and MLCK-deficient (n = 26–40) cells cultured for 5, 7, and 9 days were calculated. B, wound healing assay of control and MLCK-deficient SMCs in vitro. Phase-contrast photographs were taken at 0, 12, and 24 h after the wound was made. Cell migration distances at 12 h were measured and statistically analyzed (n = 102 and 160 respective to CTR and KO). C, consecutive photographs for protruding SMCs were taken within the period of 0–30 min. The extended distance of the leading edge was measured (n = 25 and 30 for CTR and KO, respectively) and statistically analyzed. D, 13 control and 8 MLCK-deficient SMCs were randomly selected for cell migration trace analysis as shown in the left panel. The accumulated migrated distance (middle) and average migrating velocity (right) were measured and statistically analyzed. E, the adhesion properties of control and MLCK-deficient SMCs cultured for 7 days were examined by fixing the cells in situ and co-staining them with anti-SMA (green) and anti-FAK antibodies (blue). Scale bars are 200 μm (A, B) and 20 μm (C, D, E).