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. 2017 Jan 17;127(2):635–650. doi: 10.1172/JCI89486

Figure 5. BCL-W expression is regulated by MYC through transcriptional upregulation of the miR-15 family.

Figure 5

(A–C) A luciferase reporter containing the 3′-UTR of Bcl-w with a WT or mutated (Mut) miR-15 family binding site was transfected into p53–/– MEFs expressing MYCER (A), MYCΔMBII-ER (B), or MYCV394D-ER (C). Luciferase activity was measured in triplicate 48 hours after the addition of vehicle control (EtOH) or 4-OHT. A luciferase reporter containing either the WT or mutated miR-15 family binding site of the 3′-UTR of Bcl-2 served as a positive control for A–C. A β-gal reporter was cotransfected for normalization purposes. (C and D) miR-15 family binding sites in the 3′-UTR of Bcl-w and Bcl-2 were blocked with site-specific small molecules (Target Protectors [TP]) 24 hours prior to MYCV394D-ER activation. Following the addition of 4-OHT, luciferase activity (C) and protein levels (D) were assessed in the presence or absence of the Bcl-w and Bcl-2 Target Protectors. Data shown are representative of 2 to 4 independent experiments. Error bars indicate the SD. *P = 0.0017 (A), by t test; *P < 0.0001 (B and C), by 1-way ANOVA.