Fig. 4

miR-690 directly targets p65. a Schematic representing miR-690 target sites in p65 3′UTR and base-pairing of miR-690 sequences with the 3′UTR. b Schematic diagram illustrating the design of luciferase reporter with p65 3′UTR. The sequences of miR-690 and miR-690-Mut are also shown. The underlined sequences represent wild-type and mutated seed sequences of miR-690. Rluc Renilla luciferase, Luc Firefly luciferase, pA polyadenylation signal. c miR control, miR-690, or miR-690-mut was co-transfected with psiCHECK-2-p65 3′UTR reporter into C2C12/Runx2^Dox cells. After 48 h, cells were harvested and luciferase activities were measured. Luciferase activity = (Renilla/Firefly) ratio. Renilla, raw renilla luciferase activity; Firefly, firefly luciferase activity. Data are presented as mean ± SD (n = 3). *P < 0.05 compared with miR control sample. d, e Relative mRNA level of p65 (d) and protein level of p65 (e) in miR control- or miR-690-transfected cells