(A) HeLa cells stably expressing full length N-terminal GFP-tagged PRR14 were isolated. Cells were fixed, stained with anti-GFP and anti-Lamin A/C antibodies, and imaged using confocal microscopy. Merge 1 excludes the DAPI signal. Images are representative of the highly uniform behavior of the GFP-tagged PRR14 protein.
(B) Cells were prepared similarly to Panel A, except that the anti-LBR staining was compared to GFP-tagged PRR14.
(C) (Left) HeLa cells stably expressing both full length N-terminal GFP-tagged PRR14 and mRFP-tagged HP1α were isolated. Live cell confocal imaging is shown. (Right) HeLa cells stably expressing PRR14 N-GFP 1–135 were transiently transfected with an expression plasmid encoding mRFP-HP1α, and live cell confocal imaging is shown. Arrows indicate lack of PRR14 1–135 and HP1α colocalization only at the spindle midzone and midbody. See Figure S3.