Figure 2.

Z.2.2 localizes to the nucleus and is partially incorporated into chromatin. (A) Fluorescence imaging of stably transfected HeLa Kyoto cells shows nuclear localization of all GFP-H2A variants (middle). DNA was counterstained with DAPI (top). Overlay of both channels in color is shown at the bottom (Merge; GFP: green, DAPI: blue). Scale bar = 5 µm. (B) Deconvolved images of metaphase spreads of HeLa Kyoto cells stably expressing GFP-H2A variants (middle). Merged images in color are shown below (GFP: green; DAPI: blue). Scale bar = 10 µm. (C) Chromatin from HeLa Kyoto cells stably expressing GFP-Z.2.2 was digested with MNase followed by a purification of mononucleosomes using sucrose gradient centrifugation. Isolated DNA from subsequent sucrose gradient fractions was analyzed by agarose gel electrophoresis (left). Fractions containing pure mononucleosomes (marked with asterisk) were combined and analyzed by IB (right) using αGFP antibody for the presence of GFP-Z.2.2 (top), and αH3 (bottom).