Figure 2 | Oncogene

Figure 2

From: MicroRNA-520/373 family functions as a tumor suppressor in estrogen receptor negative breast cancer by targeting NF-κB and TGF-β signaling pathways

Figure 2

miR-520/373 family directly regulates expression of the RelA/p65 NF-κB subunit in normal breast and cancer cells. (a) RELA mRNA levels in MDA-MB-231 (***P<0.001) or MCF10A (**P<0.01) cells transfected with miR-520/373 family mimics compared with control as determined by qRT–PCR. (b) RelA/p65 protein levels after transfection of MDA-MB-231 or MCF10A cells with miR-520c, miR-373 mimics, RELA knockdown compared with control-transfected cells. After 2 days, MDA-MB-231 cells were treated with 20 ng/ml TNF-α for 10 min and total cell lysates were analyzed by western blot. (c) Schematic representation of the gene structure of RELA with the predicted target sites for miR-520/373 family members. (d) Four different psiCHECK2 reporter vectors were constructed carrying the wild-type, single or double mutated RELA 3′UTR, as indicated. (e) Luciferase reporter assay of MCF-7 cells transfected with wild-type or mutated 3′UTRs of RELA mRNA and miR-520/373 mimics or negative control. Experiments were performed with four replicates and data are shown as mean±s.d.

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