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. 2017 Jul 26;8(41):70142–70155. doi: 10.18632/oncotarget.19594

Figure 2. Dynamic interaction between ORC2 and PIAS4 or SENP2.

Figure 2

(A) U2OS cells were treated with nocodazole for 24 hours and then released into fresh medium for indicated times. Immunoprecipitation was performed with ORC2 antibody and immunoprecipitates were blotted with ORC2 or PIAS4 antibody. Asterisks are indicated as non-specific bands and arrows are indicated as SUMOylated ORC2, non-SUMOylated ORC2, or IgG. (B) U2OS cells were treated with RO3306 and immunostaining was performed with PIAS4 antibody (green) and CENP-A antibody (red). Nuclei were stained with DAPI (blue). Scale bar: 10uM. (C) U2OS cells were treated with nocodazole for 24 hours and then released into fresh medium for indicated times as in (A). Immunoprecipitation was performed with ORC2 antibody and immunoprecipitates were blotted with ORC2 or SENP2 antibody. Asterisks are indicated as non-specific bands and arrows are indicated as SUMOylated ORC2, non-SUMOylated ORC2, or IgG. (D) U2OS cells were immunostained with SENP2 antibody. SENP2 staining (green) at different stages of mitosis was observed by microscope. Nuclei were stained with DAPI (blue). Scale bar: 10uM. (E) U2OS cells were transiently transfected with indicated plasmids. Cells were immunostained with HA antibody (green) and ORC2 antibody (red). Nuclei were stained with DAPI (blue). Scale bar: 10uM.