Fig. 3.

VIM-AS1 transcript knockdown results in VIM silencing with an effect on promoter CGI methylation. SW480 cells were transduced with lentiviral plasmids overexpressing control shRNA (scr) or shRNAs against VIM-AS1 RNA (sh2, sh3). (A) RT-qPCR analysis of VIM and VIM-AS1 RNAs. (B) Western blot to measure vimentin protein levels in the same transduced cells. (C) Immunofluorescence detection of endogenous vimentin. (D) LNA-based antisense oligonucleotides gapmers (ASOs) targeting intron 1 (ASO1) or exon 1 (ASO2) of VIM-AS1 transcript were transfected into SW480 cells and expression levels measured by RT-qPCR. (E) Bisulfite sequencing of regions 1 and 2 within VIM promoter CGI.