Rapamycin induced mRNA and protein expression of KLF2 in HUVECs with thrombin.

<p>After stimulation with thrombin for 4 hours, rapamycin was added to HUVECs at concentrations of 2, 20, 200 and 2000 ng/ml for 24 and 48 hours. KLF2 was assessed by real-time PCR, western blot and immunofluorescence assays. Each bar represented the mean±SD (n = 6). Quantitative data for the PCR and western studies were shown graphically. A, rapamycin induced mRNA expression of KLF2 for 24 hours. B, rapamycin induced protein expression of KLF2 for 24 hours. C, rapamycin induced mRNA expression of KLF2 for 48 hours. D, rapamycin induced protein expression of KLF2 for 48 hours. Rapamycin enhanced the expression of KLF2 compared to stimulation with thrombin alone when the blood concentration of rapamycin was higher than 20 ng/ml (*p<0.05, **p<0.01 for rapamycin + thrombin vs thrombin alone). E, immunofluorescence staining of KLF2 in the nuclei of HUVEC, controls or treated with rapamycin. KLF2 was shown in red (left column). DAPI was used to stain cell nuclei (center column). The merged image was shown in the right column.