Fig. 3: RACK1 regulates β-catenin stability through the ubiquitin-proteasome system. | Cell Death & Disease

Fig. 3: RACK1 regulates β-catenin stability through the ubiquitin-proteasome system.

From: RACK1 facilitates breast cancer progression by competitively inhibiting the binding of β-catenin to PSMD2 and enhancing the stability of β-catenin

Fig. 3

A qRT-PCR analysis of the β-catenin mRNA expression in control and RACK1-silenced breast cancer cells. B Western blotting analysis of β-catenin expression in control and RACK1-silenced breast cancer cells treated with cycloheximide (CHX). Gray scale analysis was performed using Image J software and the grayscale values of each band were normalized to the mean value of each group at the 0 hour time point. C Western blotting analysis of β-catenin expression in control and RACK1-silenced breast cancer cells treated with 10 μM of MG132. The relative expression values of the proteins were normalized to those of the control group at the 0 hour time point. D Knockdown of RACK1 resulted in increased levels of ubiquitinated β-catenin in two breast cancer cells. Control and RACK1-silenced cells were transfected with HA-ubiquitin and then treated with or without the proteasome inhibitor MG132. Afterwards, cell lysates were immunoprecipitated with anti-β-catenin antibodies and analyzed by Western blotting using anti-HA-ubiquitin antibodies. E Western blotting analysis of β-catenin expression in control and RACK1-silenced breast cancer cells treated with 50 μM of chloroquine (CQ). The relative expression values of the proteins were normalized to those of the control at the 0 hour time point. All data are expressed as the mean ± SD; ****p < 0.0001 and ns p > 0.05 versus control, N = 3.

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