Fig. 3

Cholesterol depletion induces changes in cell morphology and in the distribution of actin filaments. MDA-MB 231 cells were grown for 24 h, scratched wound lines were created and cultures were treated with MbCD (b, d, f, h). Some cells were not treated (control, a, c, e, g). Cells were analyzed 2 h after MbCD treatment by staining with the F-actin probe Texas red-Phalloidin (red, a, b, e, f) and the nuclear stain Dapi (blue, a, b, e, f) and by phase contrast microscopy (c, d, g, h). Merged images shown with Phalloidin/Dapi labeling (a, b, e, f) are the same microscopic fields as the ones shown under phase contrast microscopy (c, d, g, h). While the scratched area can be better appreciated on the low magnification (a-d), the higher magnification images (e-h) illustrate more clearly the overall cell morphology. Note that control cells (a, c, e, g) display many ruffled membranes and lamellipodia whereas in cholesterol depleted cells (b, d, f, h) membrane protusions were less prominent and most of the cells are spindle-shaped. Scale bar in A represent 50 μm in (a-d), and scale bar in E represents 50 μm in (e-h)