Fig. 1.

ADK deficiency suppresses endothelial inflammation in vitro. a Western blot detection of ADK in HUVECs treated with TNF-α at 10 ng/ml (n = 4). b Quantification of intracellular adenosine by HPLC in HUVECs exposed to TNF-α at 10 ng/ml (n = 4). c Representative images of ADK immunofluorescent (IF, green) staining on aortic endothelium (areas indicated with CD31 staining, red) of WT mice treated with vehicle or TNF-α (10 ng per gram mouse body weight for 5 h). L indicates luminal area (scale bar, 50 µm; n = 5–6 mice per group). d Heat map of inflammation-related genes in ADK KD or Ctrl HUVECs (n = 3). e RT-PCR analysis of mRNA levels of adhesion molecules in TNF-α (10 ng/ml for 2 h)-treated ADK KD or Ctrl HUVECs (n = 3). f Western blot detection of adhesion molecules in ADK KD or Ctrl HUVECs treated with TNF-α at 10 ng/ml for 3 or 6 h. Images are representative of four independent experiments. g Quantification of adhesion molecular expression in ADK KD or Ctrl HUVECs treated with TNF-α at 10 ng/ml for 6 h (n = 4). h Representative images and quantification of monocyte adhesion on TNF-α (10 ng/ml for 4 h)-treated ADK KD or Ctrl HUVECs (scale bar, 100 µm; n = 4). i Western blot detection and densitometric quantification of adhesion molecule expression in 10 ng/ml TNF-α-treated MAECs isolated from ADK^WT or ADK^VEC-KO mice (n = 4). For all bar graphs, data are the mean ± SEM, *P < 0.05 and **P < 0.01 (one-way ANOVA with Tukey’s post hoc test for a, b; unpaired, two-tailed Student’s t-test for e, i)