Fig. 2: Estrogen increases USP35 expression via decreasing miR-140-3p and miR-26a-5p levels in ER + breast cancer cells. | Cell Death & Disease

Fig. 2: Estrogen increases USP35 expression via decreasing miR-140-3p and miR-26a-5p levels in ER + breast cancer cells.

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

Fig. 2

a E2 increases USP35 protein level. MCF-7, ZR-75-1, and T-47D cells were treated with 10 nM E2 and immunoblotted with the indicated antibodies. b E2 decreases miR-26a-5p and miR-140-3p levels. MCF-7 cells were treated with vehicle or E2 (10 nM) and subjected to qRT-PCR analysis. ***, p < 0.001. c miR-140-3p and miR-26a-5p can reduce E2-enhanced USP35 protein levels. MCF-7 cells were transfected with scramble negative control RNA (NC), the indicated miRNA mimics, respectively, and treated with E2 for 24 h before being subjected to immunoblotting. d USP35 3′UTR contains the predicted miR-140-3p target sites in positions 61–66 and 738–744 (top left) and miR-26a-5p target site in position 228–234 (bottom left). MCF-7 cells were co-transfected with NC, miR-140-3p, or miR-26a-5p mimics respectively, together with USP35-3′UTR-WT-luc or USP35-3′UTR-mut-luc plasmids. The top and bottom right panels contain the USP35-3′UTR-mut-luc plasmids in which the miR-140-3p and miR-26a-5p target sites were mutated respectively. The normalized relative luciferase activities were shown on the right panels. ns not significant; ***, p < 0.001.

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