Extracellular vesicles with ubiquitinated adenosine A 2A receptor in plasma of patients with coronary artery disease

Figure 1. Presence of ubiquitinated A2AR in EV from plasma of CAD patients. Western blot of EV from plasma of four CAD patients was splitted in two parts and separately revealed using anti‐A2AR (upper part) and anti‐CD9 (lower part) primary antibody (A). Western blots of EV from four CAD patients were revealed using (from top to bottom) anti‐CD63, anti‐A2AR and anti‐ubiquitin primary antibody (B). Molecular weight markers are on the left. To compare, blots obtained with each primary antibody are mounted on the figure and are representative of triplicates

Figure 2. Comparison of EV and cell expression of A2AR. Western blots of EV (upper) and PBMC (lower) from eight CAD patients and two healthy individuals (A). Western blots of EV (upper) and CEM T cells (lower) from cells cultured in three conditions: Control, Hypoxia, Hypoxia + HCy (B). Blots were revealed using anti‐A2AR primary antibody. Molecular weight markers are on the left. To compare, blots are mounted on the figure and are representative of triplicates. Blotted bands were quantified, and results are given in % of total pixels. At the bottom of the figure, CEM cell viability after 24 h culture in the three conditions is given in absorbance at 550 nm (mean ± SD)

Figure 3. Relationship between A2AR export in EV and hyperhomocysteinemia (HHCy). Western blots of EV from six healthy individuals (upper) and six CAD patients with moderate hyperhomocysteinemia (lower). Blots were revealed using anti‐A2AR primary antibody. Molecular weight markers are on the left. Blots are representative of triplicates. Blotted bands were quantified and results are given in % of total pixels (A). Scatter plot to display the correspondence of A2AR levels in EV of CAD patients (% pixels) with their HCy levels (µmol/L) given in Table 1. Each point represents a patient (B)