Fig. 3

iSINEs can repress reporter gene expression and RNA levels experimentally. The iSINE-containing 3′ UTRs of a the Nicn1 and b Inadl genes were inserted downstream of the firefly luciferase ORF. As controls, one of the Alu elements was flipped to make a duplicated SINE (dSINE) and as a second control one of the ALUs was removed (1SINE). Arrows show SINE orientation and absolute distances are indicated. Reporter genes harboring different SINEs derived from c the Nicn1 or d the Inadl 3′ UTRs were transfected into U2OS cells and gene expression was determined using a dual luciferase assay (DLA). Total RNA was extracted from transfected cells with e the Nicn1 or f the Inadl constructs and mRNA levels were measured using reverse transcription followed by quantitative PCR of total cDNA. Standard deviation is indicated by error bars. Asterisks indicate p values calculated with Student’s t-test: *p < 0.05, **p < 0.005, ***p < 0.0005