Figure 3

The 3’UTR of HIF3α contains putative miR-210-3p and miR-485-5p binding sites and is expressed in sarcoma cell lines. (A) Schematic representation of the 3’UTR of the HIF3α transcript variants 003 and 201 (http://www.ensembl.org, release 72). Depicted are predicted target sites of miR-210-3p (open box) and miR-485-5p (black boxes). (B) Arrows indicate location of primers used to detect the presence of HIF3α 3’UTR transcripts. A 212 bp and 313 bp amplification product indicate expression of the 3’UTR of HIF3α transcription variants 003 and 201. (C) Lines designate the HIF3α 3’UTR fragments (HIF3α-short, HIF3α-long) which were cloned into the psiCHECK-2 luciferase reporter to verify regulation of HIF3α by miRNAs. (D) End-point RT-PCR was used to determine the presence of HIF3α 3’UTR transcripts and their induction upon hypoxia. Depicted are EtBr stained PCR amplification fragments of 212 and 313 bp derived from HIF3α 3’UTR cDNA and an amplified 336 bp HPRT fragment as input control. The results obtained with five representative cell lines are shown.