Figure 4.

Effect of cholesterol repletion on cell morphology, Akt activation, cell viability, and raft/caveolae distribution. A: Serum-starved A431 cells were treated either without or with 5 mmol/L MBCD or 1 mmol/L cholesterol for 1 hour. Some MBCD-treated cells were washed with medium, and then cells were incubated without or with 1 mmol/L cholesterol for 1 hour. Cells were then stained with Texas Red-labeled phalloidin to visualize actin. B: Serum-starved A431 cells were treated as described in A, and then cell lysates were subjected to immunoblotting analysis using anti-phospho-Akt antibodies. C: Serum-starved A431 cells were treated as described in A, and cell viability was measured by MTS assay. Values represent the means ± SD of three independent experiments. D: Serum-starved A431 cells were treated as described in B, and cells were stained with filipin and Alexa Fluo568-CTXB and then subjected to confocal microscopy analysis. Similar results were obtained in three different experiments.