Figure 1. Cytoplasmic Distribution and Relocalization of ZBP1

(A) Cells were fixed and stained with anti-ZBP1 antiserum (red), which was digitally superimposed on the Nomarski image and nuclear staining (DAPI, blue).

(B) Cells extracted with Triton X-100 for 30 s before treatment described in (A).

(C) Colocalization of endogenous (left) and GFP-fused (right) ZBP1 (green) and ß-actin mRNA (red) in chicken embryo fibroblasts. Left: cells were stained with anti-ZBP1 serum and were subsequently hybridized with ß-actin mRNA fluorescent oligonucleotide probes. Right: cells were transfected with the ZBP1-GFP fusion, and green cells were visualized by deconvolution microscopy after fixation and in situ hybridization. The yellow staining in the protrusions demonstrates RNA and protein colocalization. The scale bar represents 10 µm.

(D) ZBP1-GFP particles moving in living cells and localizing to a dynamic, moving lamella. Panels are shown at approximately 100-s intervals. The scale bar represents 10 µm. See Movies 1–11 in the Supplementary Material.