Figure 5.

USP1 inhibition by ML323 suppresses OS growth in vivo. A-D. Subcutaneous xenograft tumorigenesis model was established and randomized to three groups for intraperitoneal injection of PBS (negative control) or ML323 (5 and 10 mg/Kg) every two days as indicated. The nude mice were sacrificed three weeks after intraperitoneal injection and the tumors were isolated from the nude mice and arrange neatly according to each groups (A). The tumor volume and weight of nude mice were measured every two days and arranged into line charts (B). Immunohistochemistry (IHC) and Western blot assay revealed the relative content of TAZ, C-Myc, Runx2, Cyr61 and N-cadherin in tumors from different groups (C and D). E-H. A tail vein metastasis model was established and treated as mentioned. Tumor metastasis of each nude mice were imaged by an in vivo bioluminescence imaging system on day7, day14 and day21, respectively (E). The nude mice were euthanatized on day21 and the lungs were harvested for photograph and H&E staining (F, G and H). I, J. An orthotopic xenograft model was established as mentioned and intraperitoneal administration with PBS (negative control) or ML323 (5 and 10 mg/Kg) every two days. Then the nude mice were euthanatized three weeks after intraperitoneal injection and the tibia was preserved for micro-CT scanning. Data represents the means ± SD. The images and data presented were acquired from and represented three independent experiments. P*< 0.05, P** < 0.01, P*** < 0.001 in comparison with the control group.