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

Figure 1. Cell-cycle regulated ORC2 SUMOylation by PIAS4 and SENP2.

Figure 1

(A) U2OS cells were transfected with indicated plasmids. Talon beads pull-down was performed and precipitates were blotted with anti-FLAG antibody. Whole cell lysates (WCL) were blotted with HA antibody. (B) U2OS cells were transfected with indicated plasmids. Talon beads pull-down was performed and precipitates were blotted with anti-FLAG antibody. Whole cell lysates (WCL) were blotted with HA antibody. (C) U2OS cells were transfected with the indicated siRNA or control siRNA and treated with nocodazole for 24 hours. Cells were fractionated and the chromatin fraction was subjected to immunoprecipitation with ORC2 antibody. Immunoprecipitates were blotted with ORC2 or SUMO2 antibody. (D) U2OS cells were transfected with the indicated siRNA or control siRNA and treated with nocodazole for 24 hours, and then were released into fresh media for 2 hours. Cells were fractionated and the chromatin fraction was subjected to immunoprecipitation with ORC2 antibody. Immunoprecipitates were blotted with ORC2 or SUMO2 antibody. In all the figures, asterisks are indicated as non-specific bands. Arrows are indicated as SUMOylated ORC2, non-SUMOylated ORC2, or IgG.