Fig. 3

Effects of simvastatin and geranylgeranyl pyrophosphate (GGPP) on localization of di-phosphorylation of myosin light chain (ppMLC) and actin bundle formation in porcine aortic endothelial cells (PAECs). a Representative merged images of confocal microphotographs show the triple fluorescence staining of either F-actin/VE-cadherin (VE-Cad)/nuclei (DAPI) or ppMLC/VE-Cad/DAPI in PAECs in the indicated colors. The extracted images of ppMLC and actin (both in a green channel) in a gray scale are shown below the corresponding merged images. The readings of the fluorescence intensity were indicated at a lower right corner of each image. b Summaries (n = 3) of quantitative analysis of fluorescence intensities of ppMLC and actin. The data are expressed as the mean ± SEM. *P < 0.05 according to an ANOVA followed by Fisher post hoc test. PAECs were either pretreated or left untreated with 10 µM simvastatin for 16 h with and without co-treatment with 3 µM GGPP (n = 3). The images were obtained 3 min after stimulation with 1 unit/mL thrombin. The cells were challenged to thrombin, in the presence of simvastatin and GGPP, when pretreated. Control indicates images of cells without any treatment. Thrombin induced peripheral accumulation of ppMLC and actin bundles in close proximity to VE-cadherin. Simvastatin inhibited the thrombin-induced peripheral accumulation of ppMLC and actin bundles, while co-treatment with GGPP prevented the simvastatin effects. The results obtained in the two additional sets of the experiments are shown in Additional file 1: Figure S3