Journal: Pharmacological research

Article Title: In vitro angiogenesis inhibition with selective compounds targeting the key glycolytic enzyme PFKFB3.

doi: 10.1016/j.phrs.2021.105592

Figure Lengend Snippet: Fig. 6. Endothelial PFKFB3 inhibition with PA compounds act on the VEGFA/VEGFR2 axis in basal and inflammatory conditions. qRT-PCR analysis revealing downregulation of angiogenic genes (a) VEGFR2, (b) VEGFA and (c) TGF-β expression in EHECs treated with PA-1 and PA-2. (d) Relative protein expression of VEGFR2 in EHECs showing reduction upon treatment with PA-1 and PA-2. (e) By flow cytometry, a marked reduction of intracellular VEGFA production was observed in HUVECs treated with PA-1. Representative histograms show reduced VEGFA–positive ECs by PA-1 (orange) as compared to control (grey). No change was observed in HUVECs treated with PA-2 (green). (f) Heatmap indicating fold changes in expression levels of genes related to glycolysis, migration, and angio genesis upon treatment with PA-1 and PA-1 together with TNF-α in EHECs (left) and HUVEC (right). Green color represents downregulation of mRNA levels. Treatment EHEC: 20 µM 3PO, PA-1 and PA-2. Treatment HUVEC: 20 µM 3PO and PA-2, 5 µM PA-1. All experiments were performed at least three times. Data are represented as mean ± SEM, *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: Cells were washed and stained with PE-conjugated VEGFa (IC2931P, R&D Systems) for 30 min at 4 ◦C.

Techniques: Inhibition, Quantitative RT-PCR, Expressing, Flow Cytometry, Control, Migration