The 3' untranslated region of human contains regulatory elements affecting transcript stability-2

Copyright information:Taken from "The 3' untranslated region of human contains regulatory elements affecting transcript stability"http://www.biomedcentral.com/1471-2199/8/111BMC Molecular Biology 2007;8():111-111.Published online 3 Dec 2007PMCID:PMC2222623.GL4.71P control construct contained no UTR sequences. The chimeric constructs were created by cloning C1, C2, C3, C4, C5 and C6 3'-UTR fragments downstream of the Renilla luciferase gene. B) Luciferase activity of the six pGL4.71P- constructs in SK-N-BE, SH-SY5Y, HEK-293 and MCF-7 cell lines. Cells, transiently co-transfected with the pGL4.71P- constructs (Renilla luciferase) and the pGL3 (Firefly luciferase) vector were harvested 24 hours post-transfection. Luciferase activity of the chimeric constructs normalized as described, is represented as a percentage of the activity observed in cells transfected with pGL4.71P (defined as 100%). Means ± s.d. luciferase values were obtained from at least four independent experiments (* p < 0.01 compared with the corresponding pGL4.71P value). C) mRNA levels of the chimeric transcripts. Total RNA was extracted from the cells harvested 24 hours post-transfection, and the reporter gene mRNA levels were analyzed by RealTime PCR. mRNA levels of the chimeric reporter gene normalized as described, are represented as a percentage of the mRNA levels observed in cells transfected with pGL4.71P (defined as 100%). Means ± s.d. luciferase mRNA values were obtained from at least three independent experiments (* p < 0.01 compared with the corresponding pGL4.71P value).