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

Fig. 7. VHL-mediated UBE3B ubiquitination is essential for VHL in suppressing breast tumor growth and metastasis.

Fig. 7

A–C Tumor images (A), tumor growth curves (B), and tumor weight (C) of mice bearing DKO+2P2A + Myc-UBE3B-WT, DKO+2P2A + Myc-UBE3B-DM, DKO+2P2A + Myc-UBE3B-WT+Flag-VHL, and DKO+2P2A + Myc-UBE3B-DM+Flag-VHL T47D tumors (mean ± SD, n = 5). ****P < 0.0001, by 2-way ANOVA Tukey’s multiple comparisons test (B) or 1-way ANOVA Tukey’s multiple comparisons test (C). D–G Representative H&E and immunohistochemical staining of Ki-67, CC3, and endomucin in primary tumors. Scale bars: 100 μm. Ki-67-positive cell numbers (E), CC3-positive cell numbers (F), and microvessel density (G) were quantified. (mean ± SD, n = 5). ***P < 0.001, ****P < 0.0001, by 1-way ANOVA Dunnett’s multiple comparisons test. ns no significance. H, I DKO+2P2A + Myc-UBE3B-WT, DKO+2P2A + Myc-UBE3B-DM, DKO+2P2A + Myc-UBE3B-WT+Flag-VHL, and DKO+2P2A + Myc-UBE3B-DM+Flag-VHL MDA-MB-231 cells were injected into mice through tail vein. Representative H&E-stained lung sections (H) and quantification of lung metastases by qPCR (I) (mean ± SD, n = 4). ****P < 0.0001, 1-way ANOVA Tukey’s multiple comparisons test. Scale bars: 500 μm. J Immunoblots of lysates from paired normal (N) and breast cancer tissues (T). K Immunostaining of UBE3B and VHL in human breast cancer tissues. Scale bars: 25 μm.