Figure 4

Metformin reduces LD by enhancing the degradation of LD, but not by inhibiting the LD formation. (A) For the co-treatment experiment, endothelial cells were incubated without or with metformin (100 μM) concurrently with PA (200 μM) for 4 h. For the post-treatment experiments, endothelial cells were incubated without or with PA (200 μM) for 4 h, and then the cells were washed with PBS to remove PA. Then the cells were treated without or with metformin (100 μM) for another 4 h. The cells were fixed with paraformaldehyde. Cells were stained with BODIPY 493/503 (green), and nuclei were stained with Hoechst 33342 (blue). The number of LD was counted and the average number of LD per cell was calculated. (B) Five to ten microscopic fields per condition were obtained and quantified by using Image J. The quantification of LD numbers is shown in bar graphs. Data are mean ± SEM (*p < 0.05).