Figure 1

Localization and FRET between endogenous ARE binding proteins upon oxidative stress. ARE-BPs were detected via immunocytochemistry. Cells were treated with 0.5 mM sodium arsenite (SA) for 30 minutes. ARE-BPs are shown in control cells (top row), and SA treated cells, (bottom row) in each panel of images. Images in each row, from left to right, show the ARE-BP detected via: i) Cy3 labeled 2° Ab in green, ii) Cy5 or AF647 labeled 2° Ab in red, iii) Cy3-Cy5 merged image, iv) FRETc signal in thermal pseudocolor. Color matched signal intensity scales are indicated for each image. All scale bars are 10 μm in length. Panel A: HuR (Cy3) and AUF1 (Cy5). The Cy5 image intensity (bottom row) was increased to allow easier detection of AUF1 in SGs. Panel B: KSRP (Cy3) and HuR (Cy5). The gamma setting of the FRETc image (bottom row) was decreased to 0.81 to allow increased visualization of FRET interaction in SGs. Panel C: TIA-1 (Cy3) and HuR (Cy5). Panel D: KSRP (Cy3) and TIA-1 (AF647). Panel E: AUF1 (Cy3) and TIA-1 (AF647). Panel F: KSRP (Cy3) and AUF1 (Cy5). The gamma setting of the Cy5 image (bottom row) was decreased to 0.79 to allow increased visualization of AUF1 in SGs.