Figure 2

Microscopic images of cultured vascular smooth muscle cells. Cells were incubated in the absence (control, A, B, C) or presence of LL-37 (30 μg/ml) for 2 (D, E, F) or 5 (G, H, I) hours. The images show cells viewed with differential interface contrast (Nomarski) microscopy (A, D, G), stained with 4'6-diamino-2-phenyliondole dihydrochloride (DAPI), (B, E, H), or FITC-annexin V (C, F, I). Control cells have a smooth surface (A) with large evenly stained nuclei (B) and no FITC-annexin V staining of the cell membrane (C). After 2 hours' incubation with LL-37, some cells appear shrunken with a rough surface (D), smaller nuclei with fragmented chromatin (E), and a cell membrane staining with FITC-annexin V (F) indicating apoptosis. After 5 h the apoptotic changes are more marked (G, H, I; bar, 10 μm).