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  Indian J Med Microbiol
 

Figure 3: The effects of cryptotanshione (CT) on survivin and myeloid cell leukemia-1 (Mcl-1). Survivin and Mcl-1 mRNA levels were confirmed by RT-PCR in MC-3 cells and normalized to β-actin. (a) MC-3 cells were transfected with PGL3 vector control (white bar) or survivin and Mcl-1 promoter construct (black bar), treated with CT, and induction of luciferase activity was determined. (b) The effect of CT on Mcl-1 protein turnover was determined by Western blot analysis in MC-3 cells treated with the protein synthesis inhibitor, cycloheximide (0.4 μg/ml) with/without CT (8 μM) for 24 h. (c) MC-3 cells were pretreated for 1 h with the proteasome inhibitor, MG-132 (40 nM) or dimethyl sulfoxide (DMSO) and CT (8 μM) or DMSO were added for 3 h. Then, Mcl-1 expression was determined by Western blot analysis. (d) All data are expressed as means ± standard deviation for triplicate experiments and significance (P < 0.05) compared with DMSO-treated cells is indicated (*)

Figure 3: The effects of cryptotanshione (CT) on survivin and myeloid cell leukemia-1 (Mcl-1). Survivin and Mcl-1 mRNA levels were confirmed by RT-PCR in MC-3 cells and normalized to β-actin. (a) MC-3 cells were transfected with PGL3 vector control (white bar) or survivin and Mcl-1 promoter construct (black bar), treated with CT, and induction of luciferase activity was determined. (b) The effect of CT on Mcl-1 protein turnover was determined by Western blot analysis in MC-3 cells treated with the protein synthesis inhibitor, cycloheximide (0.4 μg/ml) with/without CT (8 μM) for 24 h. (c) MC-3 cells were pretreated for 1 h with the proteasome inhibitor, MG-132 (40 nM) or dimethyl sulfoxide (DMSO) and CT (8 μM) or DMSO were added for 3 h. Then, Mcl-1 expression was determined by Western blot analysis. (d) All data are expressed as means ± standard deviation for triplicate experiments and significance (<i>P</i> < 0.05) compared with DMSO-treated cells is indicated (*)