VIM-AS1 is a nuclear, polyadenylated transcript running head-to-head with VIM transcript. (A, Upper) Intronic/exonic organization of vimentin (VIM) and its antisense VIM-AS1 transcripts. Coordinates are given relative to the canonical VIM TSS and the UCSC Gene data bank (uc001iot.2) for VIM-AS1 (release hg19). (A, Lower) End-point RT-PCR from normal colon mucosa total RNA with strand-specific primers. Reverse transcription was carried out either with specific reverse primers (“R,” lanes 1–3) or with forward primers (“F,” lanes 4–6). (B, Upper) VIM-AS1 RNA transcript is polyadenylated. (B, Lower) polyA+/polyA- partition of total RNA from SW480 cells analyzed by RT-qPCR. (C) Nuclear/cytoplasmic fractionation of SW480 cells, analyzed by RT-qPCR and Western blot to assess fraction purity.