Skip to main content
. 2017 Jul 17;12(7):e0180659. doi: 10.1371/journal.pone.0180659

Fig 7. Promoter and methylation analysis.

Fig 7

A) The MSTD LTR is in three sections, being interrupted by two antisense Alu elements. Part of the last Alu and the 3’ most LTR section are not shown. Long, intermediate and short sections of the MSTD LTR / Alu sequence were cloned into the pGL4 vector immediately upstream of the luciferase gene. Locations of primers are shown as bars at the top and also in S8 Fig. The LTR-IL-33 transcriptional start site (TSS) is also shown (blue arrow). (B) The “short”, “intermediate” and “long” luciferase constructs were transfected into LS513 cells and activity assessed by luciferase assay. Data from 2 independent experiments is shown. (C) Bisulfite sequencing of native and MSTD-LTR IL-33 promoters. The promoter regions of the LTR-IL-33-expressing cell lines HT115 and LS513, and the native IL-33-expressing cell line HUVEC were subjected to bisulphite analysis where CpGs were available. Filled circles represent methylated CpGs, empty circles represent unmethylated CpGs and each row represents an independent clone. The locations of transcription start sites (TSS) are shown with green arrows.