Figure 2

Localization of Rev and Staufen-2 (hStau-2). A) hStau-2 localizes in the cytoplasm in the absence of Rev: HEK293T cells were transfected with Staufen-2-59-CMV vector and stained with goat anti-Staufen-2, Rabbit anti-goat Alexa Fluor 568 antibody and nuclear staining by DAPI. B) hStau-2 co-localizes with Rev in both the nucleoli and the cytoplasm: HEK293T cells were transfected with Rev-EGFP and Staufen-2-59-CMV vector. Localization of GFP-tagged Rev was determined by green fluorescence, hStau-2 was determined by goat anti-Staufen-2, Rabbit anti-goat Alexa Fluor 568 antibody and nucleus was stained by DAPI. Merged images show co-localization of hStau-2 and Rev. C) Graphical representation of hStau-2 localization in the absence of Rev: Representative HEK293T cells overexpressing hStau-2 were (C1, C3) used for the distribution plots (C2, C4) using Huygens Essential software twin slicer tool, where blue line denotes the boundaries of the nucleus and region outside the blue line represent the cytoplasmic region. Red lines in the plots indicate hStau-2 localization. D) Graphical representation of hStau-2 localization in the presence of Rev: Representative HEK293T cells over-expressing hStau-2 and Rev-EGFP (D1, D3) were used for distribution plots (D2, D4) using Huygens Essential software twin slicer tool, where blue line denotes the boundaries of the nucleus and the region outside the blue line represent the cytoplasmic region. Green lines in the plots indicate Rev localization and Red lines indicate hStau-2 localization. The precise cells whose graphical analyses have been shown are indicated by the white arrows.