Skip to main content
. 2007 Sep 13;104(39):15484–15489. doi: 10.1073/pnas.0703037104

Fig. 3.

Fig. 3.

Target validation of candidate miRNAs and effects of IL-1α on transfected pre-miRs. (A) The Tac1 3′ UTR was cloned into the pMIR-REPORT miRNA luciferase reporter system (pMIR-R/Tac1/SG). (B) D0 cells were cotransfected with pMIR-R/Tac1/SG and candidate pre-miRs (miR-130a, miR-206, and/or miR-302a), and luciferase and β-gal activities were measured. In parallel studies, uninduced cells were transfected with pre-miR negative control. Results are presented as the mean ± SD of normalized luciferase; n = 5. Normalizations were performed with luciferase/β-gal activities in cells transfected with pMIR-R/Tac1/SG alone, arbitrarily assigning a value of 1,500. (C) D0 cells were transfected with candidate or negative control pre-miRs or left untransfected. Cells were then stimulated for 16 h with IL-1α or unstimulated, and candidate miRNA levels were determined by real-time RT-PCR. Results are presented as mean fold change ± SD; n = 5. Normalizations with 5S rRNA were arbitrarily assigned values of 1. (D) D0 cells were prepared as in C, and SP levels were quantified by ELISA. Results are presented as mean ± SD; n = 5. *, P < 0.05 vs. negative control cells; **, P < 0.05 vs. untransfected cells stimulated with IL-1α; ***, P < 0.05 vs. untransfected, unstimulated cells.