Fig. 4: USP35 regulates ERα protein level by interacting and deubiquitinating ERα. | Cell Death & Disease

Fig. 4: USP35 regulates ERα protein level by interacting and deubiquitinating ERα.

From: USP35, regulated by estrogen and AKT, promotes breast tumorigenesis by stabilizing and enhancing transcriptional activity of estrogen receptor α

Fig. 4: USP35 regulates ERα protein level by interacting and deubiquitinating ERα.The alternative text for this image may have been generated using AI.

Knockdown of USP35 inhibits (a) and USP35 overexpression enhances (b) ERα protein level in ER+ breast cancer cells. c, d USP35 promotes ERα protein stability. MCF-7 cells infected with lentivirus expressing con-sh and USP35-sh#1 (c), and MCF-7 cells stably expressing vector, USP35WT, and USP35C450A (d) were treated in the presence of cycloheximide (CHX, 10 μM) and E2 (10 nM) for the indicated times. e MG132 reverses decreased ERα expression caused by USP35 knockdown. MCF-7 cells with con-sh or USP35-sh#1 were treated with MG132 (10 μM) for 6 h. f, g USP35 interacts with ERα in breast cancer cells. MCF-7 cell lysates were immunoprecipitated with anti-USP35 antibodies and rabbit IgG (negative control) (f), or anti-ERα antibody and mouse IgG (negative control) (g), followed by immunoblotting with the indicated antibodies. h USP35 promotes ERα deubiquitination. 293T17 cells were cotransfected with HA-ubiquitin and HA-ERα together with 3 x Flag-USP35WT or 3 x Flag-USP35C450A plasmids, and treated with MG132 (10 μM) for 6 h. Cell lysates were immunoprecipitated with anti-ERα antibody and mouse IgG.

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