Knockdown of PHD-2 and its effect on the expression of angiogenic growth factors in NIH-3T3 cells. NIH-3T3 cells were incubated for 4 h with nanoformulation loaded with 30 nM non-specific siRNA (siCtrl) or siPHD-2, followed by 20 h incubation with cell culture medium without or with 20 µm DES. After another 24 h incubation in fresh cell medium, mRNA was isolated and subjected to real-time reverse transcriptase PCR analysis to determine expression of (A) prolyl hydroxylase domain-2 (PHD-2), (B) hypoxia inducible factor-1α (HIF-1α), (C) vascular endothelial growth factor (VEGF), and (D) fibroblast growth factor-2 (FGF2). The results were normalized to the mRNA expression level of ß-actin and shown relative to cells transfected with siCtrl. Data are represented as the mean ± the standard deviation for a minimum of three independent experiments. (ns = not significant p > 0.05, * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001, **** p ≤ 0.0001).