Functional CRE element in <i>YWHAZ</i> transcript variant 1c.

<p>(<b>A</b>) The proximal promoter for transcript variant 1c was cloned upstream of a luciferase reporter vector. Truncations of the promoter region were generated and cotransfected with <i>renilla</i> control vector and assayed for relative luciferase activity 24 hours later. Data are graphed as <i>firefly</i> relative to <i>renilla</i> over empty vector control and represent averages of three technical experiments +/− one standard deviation. (<b>B</b>) The sequence between and surrounding the <i>YWHAZ</i> truncations (−72 and −85) was aligned from human to <i>Tetraodon</i>. Light grey nucleotides indicate loss of conservation. The grey box containing 100% sequence conservation contains the putative CRE element. (<b>C</b>) Schematic of the sequence surrounding the CRE element and the two-nucleotide mutation generated (star) is shown. HeLa cells were transfected with one of three variant 1c constructs with or without CRE mutations engineered into them and a <i>renilla</i> control reporter plasmid. Data are graphed as <i>firefly</i> relative to <i>renilla</i> over empty vector control and represent the average of three independent experiments +/− one standard deviation. Loss of activity for each of the three mutated reporters is indicated.