Figure 4 | Cell Death & Disease

Figure 4

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

Figure 4

Cdc20 regulates cellular levels of SMAR1 through APC/C complex. (a) SMAR1 is significantly stabilized in Cdc20KD cells. Stable knockdown cells of Cdc20 was made by specific shRNAs against Cdc20 (shCdc20) in MCF7 and scramble shRNA (NS) was used as control and lysates were immunoblotted for indicated proteins. (b) Relative mRNA level of SMAR1 in Cdc20KD and NS cells. SMAR1 mRNA levels were measured using RT-qPCR assay and normalized against GAPDH mRNA. NS represents statistically not significant difference calculated by one way ANOVA test (c) Slower turnover of SMAR1 in Cdc20KD cells. NS or Cdc20KD cells were treated with cycloheximide (40 μg/ml) for indicated periods and then cells were harvested, prepared cell lysates and immunoblotted for indicated proteins. (d) Cdc20 promotes lysine 48-linked polyubiquitylation of SMAR1. NS and Cdc20KD cells were lysed and whole-cell lysates were pull-down for SMAR1 and immunoprecipitates were immunoblotted for K48-linked ubiquitin. (e) Cdc20 degrades SMAR1 through APC/C complex. NS and APC2KD cells were transfected either with vector control or HA-Cdc20 and protein lysates were immunoblotted for indicated proteins

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