LINC00901 is an m6A-modified lncRNA and its high level is associated with poor prognosis of PDAC patients. (IMAGE)
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LINC00901 is an m6A-modified lncRNA and its high level is associated with poor prognosis of PDAC patients. (A) MeRIP and lncRNA profiling identified five candidate lncRNAs in MIA PaCa-2 cells, among them LINC00901 is a top candidate. BC200 and GAPDH were used as internal controls. (B) Validation of lncRNA profiling results in both MIA PaCa-2 and AsPC-1 cells by MeRIP and qRT-PCR assay. (C) Detection of interaction of LINC00901 with m6A antibody in MIA PaCa-2 and AsPC-1 cells. In vivo expressed S1m-tagged LINC00901 or vector control (S1m) was first pulled down by streptavidin beads and then incubated with m6A antibody overnight. The signals bound to the membrane were detected by donkey anti-mouse antibody conjugated with IRDye® 680RD. HC, heavy chain; LC, light chain. (D) RNA m6A methylation level of LINC00901 as determined by EpiQuik m6A RNA Methylation Quantification Kit. In vivo expressed S1m-tagged LINC00901 or vector control (S1m) was pulled down by streptavidin beads and then the bound S1m-tagged RNA was isolated from the precipitates by phenol/chloroform extraction. The m6A level of purified RNA was measured per the manufacturer's instruction. (E, F) The data obtained from the Kaplan–Meier plotter database (http://kmplot.com/analysis/) suggests poor overall survival (OS) and relapse-free survival (RFS) with high expression levels of LINC00901, compared with those in the low expression levels. Values are mean ± S.D., n = 3, ∗∗P < 0.01; ns, no significant.
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Genes & Diseases
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