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. 2022 May 21;23(10):5774. doi: 10.3390/ijms23105774

Figure 4.

Figure 4

4PYR regulates eAdo generation and deamination during paracrine interactions between 4T1 and H5V endothelial cells. (A–C) Effect of 4PYR (100 μM) on the rate of extracellular ATP hydrolysis (A), AMP hydrolysis (B), and adenosine deamination (C) in the co-culture of H5V and 4T1 cells, compared to single-cell type cultures. Vehicle-treated cells were used as a control. Cells were separated using a filter (pore size of 1 µm) in a Transwell chamber. 4PYR was added to the upper chamber containing 4T1 cells. Enzymatic activity was measured via reversed-phase HPLC after 72 h of culture. (D) Effect of TNF (10 ng/mL) on the rate of eATP hydrolysis by H5V cells. Enzymatic activity was measured on 96-well plates via reversed-phase HPLC after 6 or 24 h of culture. Vehicle-treated cells were used as a control. Graphs represent the mean ± SEM from four independent experiments. * p < 0.05, ** p < 0.01, *** p < 0.001.