Figure 1 | Cell Death & Disease

Figure 1

From: Cdc20 directs proteasome-mediated degradation of the tumor suppressor SMAR1 in higher grades of cancer through the anaphase promoting complex

Figure 1

SMAR1 is regulated via proteasomal pathway. (a) Endogenous level of SMAR1 is stabilized upon MG132 treatment. MCF7 cells were treated with 10 μM MG132 (proteasome inhibitor) and 100 μM Chloroquine (lysosome inhibitor) for 6 h. Cells were collected, prepared whole protein extracts and immunoblotted for indicated proteins. (b) MG132 alters the turnover kinetics of SMAR1. MCF7 cells were treated with 40 μg/ml cycloheximide in the absence and presence of 10 μM MG132 and treated cells were harvested at indicated time points. Whole cell protein extracts were analyzed by immunoblotting with indicated antibodies. Expression level of each protein on immunoblot (b) was quantified densitometrically. Then, expression of SMAR1 at each time point was normalized with tubulin expression and presented graphically. (c) Suppression of SMAR1 expression by candidate genes. Candidate genes were ectopically expressed in MCF7 cells for 48 h. Cells were then collected, prepared whole cell protein extracts and analyzed by immunoblotting with indicated antibodies. (d) Cdc20 degrades SMAR1 in a dose dependent manner. MCF7 cells were transfected with increasing doses of HA-Cdc20, prepared the protein extracts and immunoblotted for indicated proteins

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