MAD1 and MAD2L1 are enriched in the nucleus in U2OS cells after HU exposure.

<p>(A) Untreated or HU treated U2OS cells stained with MAD2L1 (red) and Mab414 (NPC)(green) with DAPI (blue). (B) First panels show U2OS cells stained with MAD2L1 (red) or MAD1 (green) and counterstained with DAPI (blue) in untreated cells, with HU or with colchicine. Second panels show U2OS cells treated with colchicine and stained with CREST (green), MAD1 (red) and DAPI (blue). CREST recognizes CENP-A, -B, and—C. (C) Graph of the average ratio of nucleoplasmic MAD2L1 or MAD1 fluorescence to cytoplasmic signal in the presence and absence of HU; ***p<0.0001; n≥50; Error bars indicate SEM. (D) Untreated and HU treated U2OS cells stained with CENPA (green) and counterstained with DAPI (blue). Scale bars 10 μm.