Fig EV3. USP8 inhibitor represses cancer progression and metastasis.

• A
  Normalized TβRII mRNA expression from MDA‐MB‐231 breast cancer cells treated with USP8 inhibitor and TGF‐β (2.5 ng/ml) as indicated.
• B
  Pre‐ranked gene set enrichment analysis (GSEA) in MDA‐MB‐231 cells treated with control DMSO versus USP8 inhibitor (2 μM) for 24 h. The consensus signature of the genes correlated with various breast tumor characteristics that enriched in cells with normal USP8 activity (control) or cells with suppressed USP8 activity (USP8 inhibitor) were shown.
• C–F
  Experimental analysis in vivo: MDA‐MB‐231‐Luc cells (1 × 10⁵ per mouse) were intracardially injected into nude mice (n = 5 for each group). After 3 weeks, USP8 inhibitor (1 mg/kg) was injected via intraperitoneal injection every the other day for three weeks. BLI signals (left panel) and BLI imaging of three representive nude mice from each group at week 6 (right panel) were shown (C). Number of bone metastasis nodules (D). Percentage of TβRII⁺ crEVs in plasma samples from mice (E). Immunoblot analysis of metastatic tumor nodule in mice (F).

Data information: *P < 0.05 (two‐tailed Student's t test (A, D, E) or two‐way ANOVA (C)). Data are shown as mean ± SD (A, C, D, E).