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. 2022 Jul 31;14(15):3732. doi: 10.3390/cancers14153732

Figure 5.

Figure 5

CDC20 regulates hnRNPU ubiquitination and its interaction with CTCF. (a,b) Ubiquitination of hnRNPU by CDC20. MDA-MB-468 control, expressing CDC20 knockdown or overexpressing CDC20 cells were crosslinked by using DSS (a) or not (b) before immunoprecipitation. The cell lysate was then immunoprecipitated with an anti-hnRNPU antibody. The level of ubiquitin was measured by immunoblot. (c) The cell lysate from MDA-MB-468 control, expressing CDC20 knockdown or overexpressing CDC20 cells was immunoprecipitated with an anti-CTCF antibody. The level of CTCF, hnRNPU, RAD21, and CDC20 was measured by immunoblot. (d) The cell lysate from MDA-MB-468 control, expressing hnRNPU-WT or hnRNPU-4KR was immunoprecipitated with an anti-V5 antibody. The level of ubiquitin was then measured by immunoblot. (e) The cell lysate from MDA-MB-468 control, expressing hnRNPU-WT or hnRNPU-4KR was immunoprecipitated with an anti-CDC20 antibody. The levels of HA-hnRNPU-WT or HA-hnRNPU-4KR were then measured by immunoblot. (f) The cell lysate from MDA-MB-468 control, expressing hnRNPU-WT or hnRNPU-4KR was immunoprecipitated with an anti-CTCF antibody. The levels of HA-hnRNPU-WT or HA-hnRNPU-4KR were then measured by immunoblot.