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. 2024 Jun 24;15(6):446. doi: 10.1038/s41419-024-06844-x

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© The Author(s) 2024

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, 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 changes were made. 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/4.0/.

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Fig. 3 — A Protein levels of Flag-VHL and Myc-UBE3B in the stable MDA-MB-231 cell line expressing Flag-VHL or Myc-UBE3B. B The cell proliferation rate was measured in the indicated MDA-MB-231 cells (mean ± SD, n = 3). ****P < 0.0001, by 2-way ANOVA Tukey’s multiple comparisons test. hrs, hours. C, D Colony formation was conducted in the indicated MDA-MB-231 cells, and representative images from three experiments were shown in (C). Colony numbers (D) were quantified (mean ± SD, n = 3). ****P < 0.0001, by 1-way ANOVA Tukey’s multiple comparisons test. E, F Invasion assays were carried out in the indicated MDA-MB-231 cells, and representative images from three experiments were shown in (E). Invaded cell numbers were quantified in (F) (mean ± SD, n = 3). ****P < 0.0001, by 1-way ANOVA Tukey’s multiple comparisons test. G Protein levels of VHL and UBE3B were detected in control, VHL-KD#1, UBE3B-KD, and VHL- and UBE3B-double knockdown (VHL-KD#1 + UBE3B-KD) MDA-MB-231 cells. H Proliferation rate of control, VHL-KD#1, UBE3B-KD, and VHL-KD#1+UBE3B-KD MDA-MB-231 cells (mean ± SD, n = 3). ****P < 0.0001, by 2-way ANOVA Tukey’s multiple comparisons test. hrs, hours. I–L Colony formation (I, J) and invasion (K, L) assays were performed in control, VHL-KD#1, UBE3B-KD, and VHL-KD#1+UBE3B-KD MDA-MB-231 cells, and the representative images from three experiments were shown. Colony numbers (J) and invaded cell numbers (L) were quantified (mean ± SD, n = 3). ****P < 0.0001, by 1-way ANOVA Tukey’s multiple comparisons test. Scale bars: 100 μm. EV, empty vector.