Fig. 4.

Ang1 activates UCP2 protein expression in endothelial cells. A, EA.hy926 cells were treated with Ang1 for the times indicated and UCP2 protein detected by immunoblotting. B, UCP2 and GAPDH mRNA were detected in cell lysates by RT/PCR following Ang1 activation. C, UCP2 protein was quantified by scanning blots from three independent experiments. UCP2 protein levels were normalized to actin and data are presented as mean and SE and shown relative to UCP2 in control-treated cells (*p < 0.05, Student's ‘t’ test). D, hnRNP-K suppresses UCP2 translation. CHO cells were transfected with plasmids containing nucleotides 1-1647 of UCP2 together with a plasmid encoding either hnRNP-K-GFP or GFP as indicated. Approximately 18 h post-transfection cells were lysed and UCP2 protein detected in cell lysates by immunoblotting. Blots were stripped and re-probed for GFP to confirm expression of hnRNP-K-GFP and GFP in appropriate cell lysates. mRNA was extracted from cell lysates and UCP2 mRNA detected by RT/PCR (15 cycles).