Ang, Zhiwei Koean, Ricky Abdi Gunawan Er, Jun Zhi Lee, Li Ting Tam, John Kit Chung Guo, Huili Ding, Jeak Ling APSs conferred elevated expression in HL-60 macrophages. <p><b>(A, B and C)</b> Schematic representations of UTR-Nluc reporters with UTR mutations are shown. The AU content in the first 100 bases of the 3’ UTR of these UTR-Nluc reporter mutants are shown in line charts. Positions 1 to 91 of the 3’ UTR of TNF was inserted in between positions 285 and 286 of the <i>CXCL8</i>-3’ reporter to generate the <i>CXCL8</i>-3’::dis<i>TNF</i>-3’(1–91) mutant. The expression plasmids for these UTR-Nluc reporter mRNAs were transfected into parallel cell cultures. The resulting Nluc protein and mRNA expression levels were quantified via luciferase assay and real-time PCR, respectively, after overnight incubation. The ratio of UTR-Nluc over Cntrl-Nluc expression was then determined and presented as log<sub>2</sub> values. Each graph symbol (squares or triangles) is the result of a replicate experiment. Replicate experiments were performed on different days. <b>(D)</b> The 3’ UTR of <i>TNFAIP6</i>, <i>IFNG</i> and <i>IL2</i> contain putative APSs. <b>(E)</b> From 14 fractions spanning the entire sucrose gradients displayed in <a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1008077#pgen.1008077.g001" target="_blank">Fig 1C</a>, the levels of specific <i>TNFAIP6</i> mRNA were quantified via real-time PCR and presented as a percentage of the sum of all fractions. Replicate experiments were performed on different days and the data from 2 replicates are shown.</p> TAK;cell type-specific expression;CXCL 8;cell type-specific expression levels;CXCL 8- UTR-reporter;TNFAIP;CXCL 8 protein;AU base content;PKA;protein S 6;CXCL 8 polyribosome association;C-X-C;rpS 6 phosphorylation;CXCL 8 synthesis;CXCL 8 mRNA;AZD;ERK;TNF;IL;CXCL 8 translation;UTR sequences;macrophage-like HL -60 cells;APS-positive mRNAs 2019-04-10
    https://plos.figshare.com/articles/figure/APSs_conferred_elevated_expression_in_HL-60_macrophages_/7979561
10.1371/journal.pgen.1008077.g005