Figure 5
From: ERG is required for the differentiation of embryonic stem cells along the endothelial lineage
ERG knockdown in ERG shRNA treated EB. (A) Four different ERG shRNA sequences were used for transduction and expression in ES cells prior to EB differentiation. A scrambled nucleotide sequence was used as a control. (B) Evaluation of ERG expression by Q-RT-PCR in EBs at day 8 using lentivirally delivered control, and ERG shRNAs (lentivirally delivered sequences #1-4 were used to generate ERGshRNA ES cell clones #1-4 respectively). Expression levels are shown as a percentage of control shRNA treated cells Error bars indicate means +/- S.D. (n = 4). (C) Evaluation of ERG expression in differentiating control and ERG shRNA treated ES cells by Western blot analysis. (D) Immunohistochemical analysis of ERG (green), VE-cadherin (red), and nuclear staining DAPI (blue) in EBs from control and ERG shRNA clones # 3 and 4 at day 10 and day 8 (# 3) The images were obtained using Leica fluorescence microscope at 40× magnification. Scale bar represents 100 μm. (E). Evaluation of VEGF-R2 expression in ERG shRNA (#3 and #4), or control shRNA treated ES cells by flow cytometry at 3 and 6 days after differentiation.