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. 2003 Apr;23(7):2316–2328. doi: 10.1128/MCB.23.7.2316-2328.2003

FIG. 7.

FIG. 7.

The 3′UTR of TNF-α mRNA confers translational repression in a pVHL-dependent manner. (A) pGL3-luciferase (pGL3-LUC) and pGL3-luciferase containing the 3′UTR of TNF-α [pGL3-LUC+TNF-α(3′)] were transiently transfected along with control plasmid pSV-β-gal into 786-0 cells that either expressed or lacked pVHL function (VHL+ and VHL−, respectively). (B) Representative Northern blot analysis used to ascertain the expression levels of LUC+TNF-α(3′) transcript (and LUC in pGL3-LUC transfections [data not shown]) and the β-galactosidase mRNA in transfected 786-0 and UOK populations; determination of 18S signals served to monitor the quality of RNA samples. Graphs represent the relative abundance of mRNAs containing the luciferase coding region [either LUC mRNA or LUC+TNF-α(3′) mRNA] in pVHL-deficient cells (100%) compared with pVHL-expressing cells. In each transfection group [pGL3-LUC and pGL3-LUC+TNF-α(3′)], luciferase mRNA values were normalized to β-galactosidase mRNA values. (C) Luciferase activity in 786-0 and UOK cells transfected with either pGL3-LUC or pGL3-LUC+TNF-α(3′); measurements of luciferase activity were normalized to measurements of β-galactosidase activity and plotted as percent luciferase activity/galactosidase activity in pVHL-expressing cells (VHL+) relative to that measured in pVHL-deficient cells (VHL−) (100%). Data represent the means and standard errors of the means from three independent experiments.