Effects of SSW smoke on microfilaments and focal adhesion plaques. Cells were treated with 1:9 smoke dilutions and different markers were analyzed. (A, B) Rhodamine-phalloidin labeling of F-actin showed that treated fibroblasts have more F-actin staining and the stress fibers appeared thicker. (C) The increase in F-actin was confirmed by staining the cells and measuring the amount of rhodamine-phalloidin present in the cells using a fluorimeter at 550–580 nm. (D, E) Fluorescence images of cells treated with SSW smoke and labeled for the focal adhesion plaque protein, vinculin. Smoke treated cells showed an increased in focal adhesion plaque formation compare with control cells. (F) Immunoblot analysis for vinculin confirms that SSW stimulates an increase in vinculin levels. For equal loading of protein in the immunoblots, please refer to grp78 blots in Fig. 4C; the same membrane was used to reprobe for the protein shown in this figure. (G, H) Effects of SSW on cell migration. Cells were plated inside cloning rings and allowed to adhere for 3 hours to form a "ring of cells". After marking the edge of the ring, the cells were treated and allowed to migrate for 24 hours and the migrated distance was measured from the edge of the ring to the migrating front of the cells. The treated cells showed a decrease in cell migration. (I) Quantification of the extent of inhibition of cell migration by SSW smoke. Data are representative of six different points along the circle. Dashed lines in G&H demark the edge of the "circle of cells". All experiments were performed at least 3 times with different batches of primary cells. Scale bar = 20 μm.
Wong et al. BMC Cell Biology 2004 5:13 doi:10.1186/1471-2121-5-13