Figure 7. UCHL1 is a good therapeutic target.

(a–c) 293T cells transiently transfected with the 5HREp-luc reporter gene (a) and HeLa/5HRE-Luc cells (b,c) with none (b) or either pcDNA4/UCHL1 (UCHL1) or pcDNA4/myc-His A (EV) (c) were treated with (+) or without (−) the UCHL1 inhibitor, LDN57444, cultured under normoxic or hypoxic conditions for 24 h, and subjected to western blotting using the indicated antibodies and luciferase assay. n=3. (d–g) Athymic nude mice were intravenously transplanted with EMT6/EF-Luc/EV or EMT6/EF-Luc/UCHL1 cells, and treated with or without LDN57444 (0.5 mg kg⁻¹ daily) from 3 to 5 days after the transplantation. (d,e) On the indicated days after transplantation, mice were subjected to an optical in vivo imaging experiment. n=6. (f,g) The relative number of metastatic colonies to the EV and LDN57444 group was analysed in the lungs 10 days after transplantation. n=6. (h–k) Athymic nude mice were transplanted with B16F10/EF-Luc cells through the tail vein, and treated with or without YC-1 (30 mg kg⁻¹ daily) or LDN57444 (0.5 mg kg⁻¹ daily) from 3 to 5 days after transplantation. (h,i) Mice were subjected to an optical in vivo imaging experiment 7 days after transplantation. n=9. (j,k) The relative number of metastatic colonies to the control group was analysed in the lungs 10 days after transplantation. n=9. Mean±s.d. *P<0.05; NS, not significant, Student’s t-test (a–d,f,h,j).