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. Author manuscript; available in PMC: 2015 Feb 1.
Published in final edited form as: Nat Methods. 2014 Jun 15;11(8):861–867. doi: 10.1038/nmeth.2998

Figure 5.

Figure 5

dfTAT-mediated delivery improves the delivery and transcriptional output of a transcription factor. (a) Assay showing that dfTAT mediated delivery of HoxB4 and TAT-HoxB4 improves the expression of a luciferase reporter. NIH 3T3 transfected with a HoxB4-dependent luciferase reporter were incubated for 1.5 h with either HoxB4 or TAT-HoxB4 (200 nM) in presence or absence of dfTAT (3 μM). TAT-mCherry (200 nM) and dfTAT (3 μM) serve as negative controls (400,000 cells/experiment, experiments in duplicate). (b) The amount of DEAC-K9 delivered in the cytosol and nucleus of live cells can be titrated. HeLa cells were incubated with dfTAT (5 μM) and increasing amounts of DEAC-K9 (1, 2.5, 5, 10, 20 μM). The fluorescence intensity of cells displaying cytosolic release was assessed by microscopy (representative 20X images are shown using a pseudocolored colorscale: blue=lowest intensity, red=highest intensity) and by measuring to the bulk fluorescence of cell lysates (microscope: 1,000 cells/experiment, fluorometer: 300,000 cells/experiment; experiments in triplicates, average and standard deviations represented). Scale bars, 10 μm (c) Assay showing that the induction of luciferase expression by dfTAT-mediated delivery of HoxB4 can be titrated. NIH 3T3 cells were co-incubated with HoxB4 (25-200 nM) and dfTAT (3 μM) and luciferase induction was measured as described in a (400,000 cells/experiment, experiments in duplicate).