Long non-coding RNA SeT and miR-155 regulate the Tnfα gene allelic expression profile
Fig 5
SeT RNA expression in wild type and miR-155-/- BMDMs.
(A) Depiction of three putative miR-155 sequence complementarity seed sites on SeT RNA, as defined by the Targetprofiler prediction tool. (B) qRT-PCR analysis for the relative SeT RNA expression normalized with Hprt1 mRNA levels in wild type and miR-155-/- mice. (C) Detection of nascent SeT RNA under non-denatured conditions in conventional RNA FISH z-stack images before and after LPS induction of wild type and miR-155-/- macrophages (scale bar 4 μm). (D) Single confocal z-stack RNA-DNA FISH images portraying the nascent mono- and biallelic expression of SeT RNA in naïve and one hour LPS-stimulated BMDMs of wild type and miR-155-/- mice (scale bar 2 μm). The corresponding graph showing the percentage of total cells expressing SeT RNA in either a mono- or bi-allelic manner was based on measurements performed in two independent experiments (sample size, WT: 508 nuclei, miR-155-/-: 414 nuclei). (E) RNA FISH performed in naïve and LPS-stimulated BMDMs depicting the cytoplasmic detection of SeT RNA in wild type and miR-155-/- BMDMs with the use of SeT biotinylated riboprobes under denatured conditions (scale bar 4 μm). Arrows indicate enlarged speckles of accumulated RNA in the cytoplasm.