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. 2024 Sep 10;7:1112. doi: 10.1038/s42003-024-06820-3

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© The Author(s) 2024

Open Access This article is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License, which permits any non-commercial use, sharing, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if you modified the licensed material. You do not have permission under this licence to share adapted material derived from this article or parts of it. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by-nc-nd/4.0/.

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Fig. 6 — A, B Effects of KDM2B^F-box-LRRs/SKP1(A) or KDM2B^F-box-LRRs(KKR/D)/SKP1 (B) on condensates of BCOR^{ANK-linker-PUFD}/PCGF1^RAWUL. Before mixing with KDM2B^F-box-LRRs/SKP1 or KDM2B^F-box-LRRs(KKR/D)/SKP1, BCOR^{ANK-linker-PUFD}/PCGF1^RAWUL (100 μM) was mixed with 5 mM CaCl₂ to stimulate droplets formation. C, D Co-localization analysis of KDM2B^F-box-LRRs/SKP1 and BCOR^{ANK-linker-PUFD}/PCGF1^RAWUL in Ca²⁺ induced droplets. For (C), KDM2B^F-box-LRRs/SKP1-mCherry or KDM2B^F-box-LRRs(KKR/D)/SKP1-mCherry (10 μM) was mixed with GFP-BCOR^{ANK-linker-PUFD}/PCGF1^RAWUL (100 μM), then 5 mM CaCl₂ was added to induce droplet formation. For (D), BCOR^{ANK-linker-PUFD}/PCGF1^RAWUL (100 μM) was incubated with 5 mM CaCl₂ to stimulate droplets formation, then KDM2B^F-box-LRRs/SKP1-mCherry or KDM2B^F-box-LRRs(KKR/D)/SKP1-mCherry (10 μM) was added to the mixture immediately. KDM2B^F-box-LRRs(KKR/D) indicates that residues K1138, K1139 and R1147 on KDM2B^F-box-LRRs were mutated to Asp. E Co-localization analysis of KDM2B^F-box-LRRs/SKP1-mCherry with CaCl₂ induced BCOR^{ANK-linker-PUFD}/PCGF1^ΔN35/Ring1B droplets. KDM2B^F-box-LRRs/SKP1-mCherry (6 μM) was mixed with BCOR^{ANK-linker-PUFD}/PCGF1^ΔN35/Ring1B (60 μM), then 5 mM CaCl₂ was added to induce droplet formation. F Co-localization analysis of BCOR-mCherry mutant, PCGF1-ECFP and KDM2B-EGFP in live HeLa cells. DNA was stained by Hoechst 33258 (Blue). G Binding affinity between BCOR^{ANK-linker-PUFD} (WFY to A)/PCGF1^RAWUL and KDM2B^F-box-LRRs/SKP1 was measured using ITC. The K_d values are shown as mean ± SD for triplicate experiments. H Effect of BCOR^WFY/A mutant on the transcription of HOXB7 and HOXA9 in HEK293T cells. Empty vector was used as a negative control. shluc was used as control shRNA. ∗∗∗∗p < 0.0001. Error bars represent standard deviation from 3 individual samples.