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Fig. 2 | BMC Cancer

Fig. 2

From: Regulation of CNKSR2 protein stability by the HECT E3 ubiquitin ligase Smurf2, and its role in breast cancer progression

Fig. 2

Forced expression of wild-type or catalytically inactive Smurf2 stabilizes CNKSR2 protein. a Co-transfection of wild-type (WT) or catalytically inactive (C716A) Smurf2 increased CNKSR2 expression. Smurf2 WT or mutant (C716A), or empty vector, was transfected into HEK293 cells, respectively. 48 h post-transfection, cells were harvested and immunoprecipitated (IP) by anti-Flag antibody followed by immunoblotting (IB) with rabbit CNKSR2 and mouse Flag (for Flag-Smurf2) antibodies. b Protein levels of CNKSR2, Flag-Smurf2, and β-actin in 1% of the total cell lysate were also shown. c Status of cellular CNKSR2 ubiquitination in Smurf2WT and Smurf2C716A transfected cells. HEK293 cells were transfected with Smurf2WT, Smurf2C716A, or empty vector respectively. After 48 h, cell lysates were harvested, denatured, and subjected to anti-CNKSR2 immunoprecipitation (IP). Precipitates were then resolved by 4–12% SDS-PAGE and immunoblotted (IB) with anti-ubiquitin antibody. CNKSR2, Flag-Smurf2 and β-actin levels were also shown by immunoblotting in 1% input

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