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. 2019 Apr 5;316(6):H1468–H1479. doi: 10.1152/ajpheart.00690.2018

Fig. 3.

Fig. 3.

CYP4A/20-hydroxyeicosatetraenoic acid (20-HETE) is associated with endothelial progenitor cell (EPC) differentiation and functions. Human umbilical cord blood-derived EPCs were isolated and seeded to allow spontaneous differentiation in culture. 20-HETE synthase CYP4A11 (A) and 20-HETE production (B) were determined on days 9, 12, 14, and 21 after EPC isolation, along with fully differentiated endothelial cells (ECs) from these EPCs (on day 28), by Western blot and LC-MS/MS, respectively (mean ± SD; n = 4; *P < 0.05 vs. day 9 EPCs; #P < 0.05 vs. differentiated ECs). EPC cultures (3 × 106) were harvested on days 9, 12, 14, 21, and 28, and colony formation Hill assay was performed according to the manufacturer’s recommendations (C; mean ± SD; n = 3; *P < 0.05 vs. day 9 EPCs; #P < 0.05 vs. differentiated ECs). Representative EPC colony is shown. EPCs (3 × 106) were harvested on day 21, and colony formation Hill assay was again performed according to the manufacturer’s recommendations in the presence and absence of 10 nM 20-HETE and/or 10 nM 6,15-20-HEDGE (D; mean ± SD; n = 3; *P < 0.05 vs. control). N.D., not detectible.