Increased miR-155 levels result in reduced mRNA levels of several HIV-1 dependency factors.

<p>We isolated total RNA at: (i) 48 h post-transfection of MDMs from five different donors with either pEZX-miR-155 or pEZX-scrambled control (A); and (ii) 24 h post-stimulation of MDMs from two different donors with poly(I∶C) or LPS (B); and then performed relative quantitation of mRNAs by real-time RT-PCR of two experimentally-demonstrated miR-155 targets (SHIP1 and SOCS1), and four not previously described, potential miR-155 targets (ADAM10, TNPO3, LEDGF and Nup153), using TaqMan Gene Expression assays (Applied Biosystems). <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1002937#s3" target="_blank">Results</a> were calculated using 18S rRNA as internal control and the comparative Ct method (also known as the 2^−ΔΔCt method), and are shown as fold-change (mean ± SD from two independent quantitations) with respect to the untransfected control in (A), or as fold-change (calculated from a single experiment) with respect to unstimulated in (B).