Figure 2

Localization and FRET interactions of exogenous ARE binding proteins upon oxidative stress. Vector constructs of ARE binding proteins labeled with eYFP or eCFP were transiently transfected into DDT1-MF2 cells. Cells were treated with 0.5 mM sodium arsenite for 30 minutes. The localization of ARE-BPs is shown in control cells (top row), and sodium arsenite treated cells (bottom row), in each panel of images. Images in each row, from left to right, show the localization of an ARE-BP detected via: i) YFP in green, ii) CFP in blue, iii) merged YFP and CFP images, iv) FRETc signal in a thermal pseudocolor scale. A color matched signal intensity scale is indicated for each image. All scale bars are 10 μm in length. Cell outline as shown. Panel A: HuR-YFP and p37AUF1-CFP. Panel B: HuR-YFP and KSRP-CFP. The signal intensity of the KSRP-CFP image (bottom row) was increased to more easily view SGs. Panel C: TIA-1-YFP and HuR-CFP. Panel D: TIA-1-YFP and KSRP-CFP. Images are of live cells under DMEM. Panel E: TIA-1-YFP and p37AUF1-CFP. A very small portion of p37AUF1 is detectable in SGs, (elongated SG, lower right side of cell). Panel F: p37AUF1-YFP and KSRP-CFP.