Fig. 1
Increased expression of Prkaa1/AMPK in ECs exposed to disturbed flow. a Representative images of en face immunofluorescence staining and quantification data of pPrkaa1 (Thr172) and Prkaa1 (red) levels in the arterial endothelium of C57BL/6j mice. The endothelium was visualized by CD31 staining (Alexa Fluor-488, green), and nuclei were counterstained with DAPI (blue). Images were captured with confocal fluorescence microscopy. Scale bar: 20 µm; n = 10 mice per group. Boxes in image on the right indicate origin of regions shown in the respective rows of images (scale bar: 5 mm). b Schematic illustration of laminar flow (shear stress: 15 dyne/cm2) and oscillating flow (shear stress: ± 5 dyne/cm2, frequency: 1 Hz) systems in vitro. c Real-time PCR analysis of mRNA levels of PRKAA1, PRKAA2, PRKAB1, and PRKAG1 in HUVECs under laminar flow and oscillating flow for 24 h. n = 4. d Western-blot analysis and quantification data of protein levels of pPRKA (Thr172), PRKAA1, PRKAA2, total PRKAA, PRKAB1, and pACC (Ser 79) in HUVECs under laminar flow and oscillating flow for 24 h. β-Actin was used as a loading control. n = 5. e Western-blot analysis and quantification data of the protein levels of pPRKA and PRKAA1 in HUVECs transfected with siCtrl and siPECAM-1 under laminar flow and oscillating flow for 24 h. n = 5. f Mouse partial carotid ligation model and intimal RNA extraction steps from carotid arteries following flushing arteries with QIAzol lysis reagent. g Real-time PCR analysis of mRNA levels of Prkaa1, Prkaa2, Prkab1, and Prkag1 in ECs obtained from sham-operated right common carotid arteries and partially ligated left common carotid arteries in C57BL/6j mice. n = 9 mice per group. All data were expressed as mean ± SEM. Statistical significance was determined by unpaired Student’s t-test (for c, d, g) and one-way ANOVA followed by Bonferroni test (for a, e). *p < 0.05 was considered significant, **p < 0.01, ***p < 0.001
