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. Author manuscript; available in PMC: 2016 Sep 2.
Published in final edited form as: J Vis Exp. 2015 Sep 2;(103):10.3791/53175. doi: 10.3791/53175

Figure 2.

Figure 2

Delivery of dfTAT into the cytosol and nucleus of live cells was achieved in multiple cell lines. The cell lines HeLa, NIH 3T3, COLO 316 and HaCaT, Neuro-2a, MCH58 and HDF were incubated with 5 μM dfTAT for 1 h, washed and imaged. The fluorescence signal detected was in the cytosol and nucleus of cells (top panel: 20X objective, bottom panel: 100X objective). After imaging, cells were incubated at 37 ºC in a humidified atmosphere containing 5% CO2 for 24 h, washed and imaged again (top panel: 20X objective, bottom panel: 100X objective). The cell morphology did not change after 24 h. Cell viability is assessed by exclusion of the cell-impermeable nuclear stain SYTOX Blue at both 1h and 24 h time point. The TMR fluorescence at the 24 h time point is different to that obtained at the 1 hctime point presumably because of the intracellular degradation of the peptide. Scale bars, 20X objective: 50 μm; 100X objective: 10 μm.