Fig. 3: DUB-IN-2 inhibits the cell phenotype of TNBC cells.

A Western blot showed the expression level of USP8 protein in BT549 or MDA-MB-231 cell lines treated with DMSO or different concentrations of DUBs-IN-2 for 8 h. To ensure internal control, β-actin was utilized. B RT-qPCR results of USP8 mRNA expression levels in BT549 or MDA-MB-231 cell lines treated with DMSO or different concentrations of DUBs-IN-2 for 8 h. C To evaluate cell viability, the CCK8 assay was conducted on BT549 or MDA-MB-231 cell lines exposed to DMSO or DUB-IN-2 for 12 h at the designated time. D, E Wound healing assay was used to determine the migration ability of BT549 or MDA-MB-231 cell lines treated with DMSO or DUB-IN-2 for 12 h. The right panel shows the quantitative results of cell migration. F, G Transwell assay was utilized to measure the migration and invasion proficiency of BT549 or MDA-MB-231 cell lines treated with DMSO or DUB-IN-2 for 12 h. The right panel shows the quantitative results of cell migration and invasion. H, I EdU assay was conducted to examine the proliferation of BT549 or MDA-MB-231 cell lines treated with DMSO or DUB-IN-2 for 12 h. The right panel shows the quantitative results of cell proliferation. J, K Flow cytometry was used to determine the apoptosis level of BT549 or MDA-MB-231 cell lines treated with DMSO or DUB-IN-2 for 12 h. The right panel shows the quantitative results of apoptosis. L–N BT549 cells were subcutaneously inoculated into 4-week-old BALB/c female nude mice treated with DMSO or DUB-IN-2 (1 mg/kg). Mice were euthanized 35 days post-injection, and xenograft tumors were excised. Displayed are representative tumor images (L), tumor weight (M), and tumor volume (N). O, P IHC staining shows the expression levels of USP8, YAP, and Ki67 in xenografts treated with DUB-IN-2. The panel shows the quantitative results of Ki67. Scale bars, 100 μm (10X), 400 μm (40X). All Data are shown as mean ± SD, N = 3. *P < 0.05; **P < 0.01; ***P < 0.001.