Functional validation of a miRNA-34a-TREM2–3’UTR interaction.

<p><b>(A)</b> ribonucleotide sequence of the 299 nt TREM2-mRNA-3’-UTR is shown in the 5’-3’ direction; the 22 nt miRNA-34a-TREM2 3’UTR complementarity-interaction region is indicated by a black underline and the 8 nt TREM2-mRNA-3’-UTR seed sequence <b>5’-ACACUGCU-3’</b> is overlaid in yellow; a single arrowhead indicates the 5’ end of a poly A+ tail in the TREM2 mRNA (22 ‘A’ nt shown; the length of this poly A+ tail is variable); TREM2 mRNA sequence derived from NM_018965; TREM2 transcript is the major X1 variant (see also <a href="http://switchdb.switchgeargenomics.com/productinfo/id_801321/" target="_blank">http://switchdb.switchgeargenomics.com/productinfo/id_801321/</a>) (<b><a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0150211#pone.0150211.g003" target="_blank">Fig 3</a></b>); <b>(B)</b> TREM2-mRNA-3’UTR expression vector luciferase reporter assay (pLightSwitch-3’UTR; Cat#S801178; Switchgear Genomics, Palo Alto CA); in this vector, the entire 299 nucleotide TREM2 3’UTR was ligated into the unique Nhe1-Xho1 site; <b>(C)</b> control C8B4 murine microglial cells, 1 week in culture; phase contrast bright field microscopy 20x; C8B4 cells transfected with the TREM2-mRNA-3’UTR expression vector luciferase reporter were treated exogenously with miRNA-34a, a scrambled control miRNA-34a (miRNA-sc) or control miRNA-183; see references and text for further details [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0150211#pone.0150211.ref018" target="_blank">18</a>,<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0150211#pone.0150211.ref019" target="_blank">19</a>]; <b>(D)</b> compared to control, C8B4 cells transfected with a scrambled (<b>sc</b>) control pLightSwitch-3’UTR vector, the TREM2-mRNA-3‘UTR vector exhibited decreased luciferase signal to a mean of 0.16-fold of controls in the presence of miRNA-34a; this same vector exhibited no change in the presence of the control miRNA-34a-sc or miRNA-183; for each experiment (using different batches of MG cells) a control luciferase signal was generated and included separate appropriate controls with each analysis; in addition a control vector β-actin-3’UTR showed no significant effects on the relative luciferase signal yield after treatment with either miRNA-183 or miRNA-34a (data not shown); dashed horizontal line set to 1.0 for ease of comparison; N = 5; *<i>p</i><0.001 (ANOVA). The results suggest a physiologically relevant miRNA-34a- TREM2-mRNA-3‘UTR interaction and a miRNA-34a-mediated down-regulation of TREM2 expression in stressed MG cells. This pathogenic ineraction may be related to the down-regulation of other immune system genes by up-regulated pro-inflammatory miRNAs in the CNS [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0150211#pone.0150211.ref012" target="_blank">12</a>,<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0150211#pone.0150211.ref019" target="_blank">19</a>,<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0150211#pone.0150211.ref022" target="_blank">22</a>] and/or an impairment in cellular phagocytosis or related phagocytic signaling [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0150211#pone.0150211.ref005" target="_blank">5</a>–<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0150211#pone.0150211.ref007" target="_blank">7</a>,<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0150211#pone.0150211.ref020" target="_blank">20</a>,<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0150211#pone.0150211.ref021" target="_blank">21</a>].</p

A Nonsteroidal Novel Formulation Targeting Inflammatory and Pruritus-related Mediators Modulates Experimental Allergic Contact Dermatitis

<p><b>Article full text</b></p> <p><br></p> <p>The full text of this article can be found here<b>.</b> <a href="https://link.springer.com/article/10.1007/s13555-018-0223-8">https://link.springer.com/article/10.1007/s13555-018-0223-8</a></p><p></p> <p><br></p> <p><b>Provide enhanced content for this article</b></p> <p><br></p> <p>If you are an author of this publication and would like to provide additional enhanced content for your article then please contact <a href="http://www.medengine.com/Redeem/”mailto:[email protected]”"><b>[email protected]</b></a>.</p> <p> </p> <p>The journal offers a range of additional features designed to increase visibility and readership. All features will be thoroughly peer reviewed to ensure the content is of the highest scientific standard and all features are marked as ‘peer reviewed’ to ensure readers are aware that the content has been reviewed to the same level as the articles they are being presented alongside. Moreover, all sponsorship and disclosure information is included to provide complete transparency and adherence to good publication practices. This ensures that however the content is reached the reader has a full understanding of its origin. No fees are charged for hosting additional open access content.</p> <p><br></p> <p>Other enhanced features include, but are not limited to:</p> <p><br></p> <p>• Slide decks</p> <p>• Videos and animations</p> <p>• Audio abstracts</p> <p>• Audio slides</p