Identification, description, and expression of E7-less STING isoforms.

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Identification, description, and expression of E7-less STING isoforms. Identification, description, and expression of E7-less STING isoforms. (A) hSTING mRNA: schematic representation of mature hSTING mRNA. Eight exons are depicted with the corresponding exon starting nucleotides. Above, primers designed to amplify the hSTING coding sequence. (B) hSTING products amplified by RT-PCR from THP-1 and HeLa cells. (C) Schematic description of the different isolated hSTING mRNAs. The start codon is depicted in green, and the stop codon is depicted in red. Isoform 1 is characterized by the loss of exon 7. Isoform 2 lacks exon 4 and 7. Isoform 3 contains an unspliced intron after exon 3. (D) Amino acid alignment of wt hSTING and alternatively spliced new isoforms. Blue color indicates common sequence, and red highlights sequences corresponding to C-terminal peptides unique to each isoform. wt hSTING and E7-less isoforms were quantified by quantitative PCR using specific primers in PBLs (E), liver biopsy specimens (G), and different cell lines (F). (H) Endogenous expression of E7-less isoform 1 in THP1 cells was tested by immune precipitation using 293T cells transfected with plasmids expressing wt hSTING and E7-less isoform 1 as reference. GenBank accession numbers: hSTING isoform 1: MH201427; hSTING isoform 2: MH201428; and hSTING isoform 3: MH201429. Estefanía Rodríguez-García et al. ImmunoHorizons 2018;2:363-376 Copyright © 2018 The Authors