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Journal: Cells
Article Title: Caffeine Mitigates Adenosine-Mediated Angiogenic Properties of Choroidal Endothelial Cells Through Antagonism of A 1 Adenosine Receptor and PI3K-AKT Axis
doi: 10.3390/cells15010087
Figure Lengend Snippet: NECA promotes AKT phosphorylation in ChEC, which is dependent on PI3K but not VEGFR2 activity. ( A ). The expression levels of phosphorylated AKT, total AKT, and β-actin were detected by Western blot following incubation with DMSO (con), NECA (N), or LY294002 + NECA (Ly + N) treatment. ( B ) Relative expression of p AKT and t AKT (total AKT) was quantified using ImageJ and Prism software (* p < 0.05; ** p < 0.01; NS: not significant). NECA-mediated enhanced AKT phosphorylation was independent of VEFGR2 activity. ( C ) The expression of phosphorylated AKT, total AKT, and β-actin was detected by Western blot after DMSO (Con), NECA (N), or SU5402 + NECA (SU + N) treatment. ( D ) Relative expression of p AKT and t AKT was quantified using ImageJ and Prism software (* p < 0.05; NS: not significant, n = 5). NECA promotes the migration of ChEC, which is mitigated by incubation with the PI3K inhibitor. ( E ) Phalloidin (green) and DAPI (blue) staining were used to detect migrated ChEC following incubation with DMSO (con), NECA (N), or LY294002 + NECA (Ly + N) within Transwells. ( F ) The cells that were transferred through the Transwell were counted using ImageJ, and data were analyzed using the Prism software (* p < 0.05, n = 8).
Article Snippet: Caffeine citrate (51754-0500-1; Exela Pharma Sciences, Lenoir, NC, USA), Bz-ATP (2′(3′)-O-(4-benzoylbenzoyl) adenosine -5ʹ -triphosphate, tri(triethylammonium) salt (HY-136254), NECA (5′-N-ethylcarboxamidoadenosine, HY-103173), DPCPX (A 1 AR antagonist, HY-100937), Istradefylline (A 2A AR antagonist, HY-10888), MRS1754 (A 2B AR antagonist, HY-14121), MRS1523 (A 3 AR antagonist, HY-121119),
Techniques: Phospho-proteomics, Activity Assay, Expressing, Western Blot, Incubation, Software, Migration, Staining