Laminar flow inhibits the Hippo/YAP pathway via autophagy and SIRT1-mediated deacetylation against atherosclerosis

摘要

HUVECs were exposed to UF (12 dyn/cm ) or DF (0.5 ± 6 dyn/cm , 1 Hz) for the indicated times. Cells with static treatment (ST) were a control. After treatment, cells were underwent immunofluorescence staining with YAP (red) and DAPI (blue). Western blot analysis showed YAP expression and YAP subcellular distribution (NP, nuclear protein; CP, cytoplasmic protein) under UF and DF flow condition in HUVEC cells. Numbers under the blots were mean ± SD of three biologically independent experiments, and the first lane (0 h) was served as relative control. Analyzed the relative total YAP protein level in nuclear and cytoplasm using gray density analysis for five independent experiments. GAPDH and Lamin B were used as a loading control for cytoplasmic and nuclear fractions. HUVECs were transfected with 8 × GTIIC reporter and renilla-luc plasmids and were subjected to shear flow 48 h post transfection. Then the relative firefly/renilla luciferase activity was determined. The expression of YAP target genes and was determined by real-time PCR. Data represent mean ± SD from three independent experiments (**  P