Figure 7.

tsRNA-1599 regulates glycolytic balance in endothelial cells. (A-E) HUVECs were transfected with negative control (NC) inhibitor (30 nM), tsRNA-1599 inhibitor (30 nM), or left untreated, and the exposed to CoCl₂ (300 μmol/L) to mimic hypoxic condition for 24 h. The group cultured in normal condition was taken as Ctrl group. Seahorse analysis of glycolysis (ECAR) was conducted at 24 h following treatment (A). The concentration of reagents used in ECAR assays was as followed: glucose (10 mM), oligomycin (2 μM), and 2‐deoxyglucose (2‐DG, 100 mM). ECAR analysis was conducted in HUVECs following tsRNA-1599 inhibition (B, n = 3, *P < 0.05 vs. Ctrl group; ^#P < 0.05 between the marked group; Kruskal-Wallis's test followed by Bonferroni's post hoc test). Glucose levels in culture medium were detected following tsRNA-1599 inhibition (C, n = 5, *P < 0.05 vs. Ctrl group; ^#P < 0.05 between the marked group; Kruskal-Wallis's test followed by Bonferroni's post hoc test). NAD⁺/NADH ratio was determined in HUVECs following tsRNA-1599 inhibition and the absorbance was measured at 450 nm (D, n = 5, *P < 0.05 vs. Ctrl group; ^#P < 0.05 between the marked group; Kruskal-Wallis's test followed by Bonferroni's post hoc test). Pyruvate level was determined in HUVECs following tsRNA-1599 inhibition and the absorbance was measured at 520 nm (E, n = 4, *P < 0.05 vs. Ctrl group; ^#P < 0.05 between the marked group; One-way ANOVA followed by Bonferroni's post hoc test). (F) Representative images of choroidal explants cultured in the presence or absence of tsRNA-1599 agomir with or without 2-DG (50 mM). The sprouting potency of choroidal explants were photographed on day 4, day 5, and day 6 (Scale bar, 500 μm).