Figure 1

Nucleolar localization of BLM requires HERC2 and its E3 domain. (a,b) HeLa-shHERC2 cells, with or without Dox-mediated induction, were subjected to immunoblotting (a) or immunostain (b) with the indicated antibodies. The nuclei were counter stained with DAPI. Scale bar, 10 μm. (c) Quantifications of the nucleolar BLM positive cell from (b) (left panel), and those from HeLa cells with a different shRNA (shHERC2#2, right panel, see also Supplementary Figure S1) are shown. Error bars, S.D. of three independent experiments, each based on more than 100 cells. Statistical significances was calculated using the Student’s t-test. (d,e) Wild-type or HERC2^ΔE3/ΔE3 HCT116 cells were subjected to immunoblotting (d) or immunostaining (e) with the indicated antibodies. Antibodies HERC2 (CT) and HERC2 recognize residues 4389–4834 and 1781–1974 of HERC2, respectively. Scale bar, 10 μm. (f) Quantification of the nucleolar BLM positive cell. Error bars, S.D. of three independent experiments, each based on more than 100 cells. Statistical significances was calculated using the Student’s t-test. Full-length blots/gels are presented in Supplementary Figure S12.