Figure 3.

GW0742 and VEGF-A enhance flux through central metabolic pathways in dynamic HUVEC. (a) VEGF-A (25 ng/ml; 4 h), but not GW0742 (100 nM; 4 h), treatment significantly increased glycolytic flux. (b) No change in D-U-¹⁴C-glucose oxidation rate was detected with either treatment. (c) GW0742 (100 nM; 5 h), but not VEGF-A (25 ng/ml; 5 h), treatment significantly increased FAO. Data are means (±S.E.M) of n = 3–5; *p < 0.05 **p < 0.01 vs. untreated control as determined by one-way ANOVA followed by Tukey’s post-comparison test. (d - h) RT-qPCR showing changes in LDHA, LDHB, MCT1, CPT1A and CACT mRNA expression in tubulogenic HUVEC treated with VEGF-A (25 ng/ml; 4 h) or GW0742 (100 nM; 4 h). Data show means (±S.E.M) of n = 3–5 *p < 0.05 **p < 0.01 ***p < 0.001 as determined by one-way ANOVA followed by Tukey’s post-comparison test. (i) Relative cellular ATP level is significantly increased in sub-confluent HUVEC treated with VEGF-A but not GW0742. Data are means (±S.E.M) of n = 3 **p < 0.01 as determined by one-way ANOVA followed by Tukey’s post-comparison test. (j) RT-qPCR for metabolic genes in mouse cardiac ECs from control (Tie2-CreERT2 + Tamoxifen; n = 5) or EC-selective tamoxifen-inducible PPARβ/δ overexpressing (Tie2-CreERT2;PPARβ/δ; n = 5) animals treated with tamoxifen compared to Tie2-CreERT2;PPARβ/δ animals (n = 5) treated with vehicle. Data are means (±S.E.M); *p < 0.05 vs. Tie2-CreERT2;PPARβ/δ + vehicle and ^#p < 0.05 vs. Tie2-CreERT2 + tamoxifen; as determined by one-way ANOVA followed by Tukey’s post-comparison test. Numbers above bars indicate p-values.