5′ Unidirectional deletion analysis of the long CIRP 5′-UTR. Ten nucleotide truncated fragments of the long 5′-UTR were generated by PCR and subcloned into the pRF bicistronic vector. Subsequently, the generated constructs were transfected into NIH-3T3 cells and grown at normothermic (37°C gray) or hypothermic (32°C black) temperatures and the luciferase activity determined 24 h post-transfection. The firefly to renilla luciferase expression ratio is shown after normalization to control β-galactosidase expression. The pRF vector was used as a control. All data shown are the means of triplicate experiments ± SD. For further details see the text.
