Figure 4. CP2005 induces pVHL through targeting USP9X to inhibit tumor cells growth.

A. and B. HepG2 and PC3 cells were treated with CP2005 (2.5 μM) for indicated periods of time. After harvesting cells were immunoblotted with indicated antibodies. C. and D. HepG2 and PC3 cells were treated with vehicle control or CP2005 (2.5 μM) for 1 hour, and GLUT1 mRNA levels were determined by qPCR. The expression levels are normalized to the GAPDH mRNA level. The results represent the mean ± SEM of three independent experiments and were analyzed with the Student's t-test. ** p < 0.01; * p < 0.05 E. HepG2 cells were treated with vehicle control or CP2005 for 1 hour. 1 × 10⁴ indicated cells were then used for evaluation of ATP levels. The results represent the mean ± SEM of three independent experiments and were analyzed with the Student's t-test. *** p < 0.001. F. Colony formation assays were performed in several cancer cell lines under CP2005 treatment. 2 × 10³ indicated cancer cells were plated in 6-well plates at 2× 10³ cells per well, and treated with various concentrations of CP2005 for 1 week. Cells were fixed and stained with crystal violet.