A. HepG2 cells were treated with either DCB-3503 or CHX for the indicated time. mRNA levels of cyclin D1, survivin, β-catenin, p53, and p21 were analyzed by real-time RT-PCR using specific primers and probes and were normalized to that of β-actin. Values are depicted as mean ± S.D. from at least three separate experiments and are expressed relative to the basal mRNA level in the corresponding untreated control cells. B. mRNA level of cyclin D1 was analyzed by Northern blot under DCB-3503 treatment. Ribosomal RNA gel image on the bottom panel shows equal loading of total RNA. C. Expressions of cyclin D1, survivin, β-catenin, p53, and p21 under treatment of CHX, DCB-3503, and CHX and DCB-3503 in combination were examined by Western blot analysis in HepG2 cells. β-Actin blot was used as internal loading control. D. (a) 2 µM MG-132 reverses suppression of cyclin D1, survivin, β-catenin, p53, and p21 by DCB-3503 within 4-hour culture, (b) 2 µM MG-132 does not block cytotoxicity of DCB-3503 after 24-hour treatment. B, C, and D are representatives from at least three separate experiments. Numbers marked in C are normalized band intensity. (* Indicating non-specific band).