Figure 5. Specific microRNA mimics and inhibitors modulated the expression of Δ;Np63α via its 3′-UTR sequences upon cisplatin exposure. (A) Predicted “seed” sequences for specific miRs in the TP63 3′-UTR with the target prediction scores in parentheses. SCC-11 cells (B) and SCC-11M cells (C) were transfected with the LightSwitch_3UTR vector for the TP63 3′-UTR along with the scrambled microRNA, or mimics and inhibitors for miR-181a-5p, miR-374a-5p, miR-519-3p, miR-630 and miR-885-3p for 36 h. Cells were treated with control medium without cisplatin (Con) or medium with 10 μg/ml cisplatin (CIS) for additional 12 h and then tested for the RenSP Renilla luciferase reporter activity. Measurements (in triplicate) for the luciferase activity presented as relative units (RU). Values obtained from cells transfected with the scrambled RNA and treated with control medium were designated as 1. (D) Total lysates from the resulting SCC-11M cells from (C) treated with control medium were subsequently analyzed by immunoblotting with antibodies to Δ;Np63α and β-actin. Relative levels of Δ;Np63α normalized for β-actin levels were quantified and shown above immunoblot images. Levels of Δ;Np63α in cells with the scrambled microRNA were designated as 1.