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. 2011 Sep;85(17):8738–8751. doi: 10.1128/JVI.00798-11

Fig. 5.

Fig. 5.

Active USP is required for stabilization in trans. (a) Cells were transfected as for Fig. 4b with constructs for constitutive expression of NT3 (lanes 1, 2, and 5, 6) or NT3.C65A (lanes 3, 4 and 7, 8), together with vectors for the Dox-inducible expression of NT1 or NT1.C65A as indicated. Twenty-four hours after transfection, Dox was added to one set of duplicate transfections and cells were incubated for a further 24 h. Samples were then probed simultaneously for expression of the various species. Dox induction of NT1 resulted in expression of NT3.C65A (lanes 3 and 4), while no increase was observed after induction of NT1.C65A (cf. lanes 7 and 8). (b) Exactly as for panel a, but comparing the effect of NT2 versus NT2.C65A on NT3 and NT3.C65A levels. (c) Cellular protein subject to proteosome-mediated turnover shows little increase upon induction of NT1. Cells were transfected as for panel a, with vectors for expression of NT3.C65A or CREBHΔTMC and processed as described. Despite significant increase in NT3.C65A (lanes 1 and 2), little effect was observed for CREBHΔTMC (lanes 3 and 4).