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. 2002 Jan;22(1):245–256. doi: 10.1128/MCB.22.1.245-256.2002

FIG. 3.

FIG. 3.

Nuclear mRNA export mediated by MS2-hTap requires hNXT binding. (A) Mammalian two-hybrid analysis. This assay was performed in 293T cells with the novel pHIV/MS2/CAT indicator construct. Cells were transfected with 100 ng of pHIV/MS2/CAT, 500 ng of the indicated MS2 fusion protein expression plasmid, 500 ng of a plasmid expressing the Tat-hNXT1 fusion protein, and 50 ng of the pBC12/CMV/β-Gal internal control plasmid. The parental pBC12/CMV plasmid served as the negative control (NEG). Induced CAT enzyme activities are given as a percentage of the activity seen in the culture transfected with pHIV/MS2/CAT, pMS2-hTap, and pTat-hNXT1. An average of three experiments with the standard deviation is indicated. (B) This nuclear mRNA export assay was performed as described in the legend to Fig. 1B. Induced CAT enzyme activities are given as a percentage of the activity observed in the culture transfected with pDM128/4XMS2 plus pMS2-hTap. An average of three experiments with the standard deviation is indicated. (C) Western analysis of the level of expression of the indicated MS2-hTAP fusion proteins in transfected 293T cells.