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Deregulation of miRNAs in <i>FTO</i> knockdown cells.

By Tea Berulava (255420), Sven Rahmann (2715), Katrin Rademacher (239174), Ludgar Klein-Hitpass (696651) and Bernhard Horsthemke (116259)

Abstract

<p>Mature miRNAs showing increased (A) and decreased (B) steady state levels in <i>FTO</i> knockdown cells. Normalized RNA-seq read numbers of individual miRNAs in <i>FTO</i> knockdown and scrambled siRNA treated cells were compared. Mean ± SD of three independent experiments are depicted. For verification and further studies, qRT-PCR analyses of selected mature miRNAs (C) and primary miRNA transcripts (D) were performed. We did not use other small RNAs as a reference gene for measuring mature miRNAs levels (as suggested by Life Technologies), since depletion of <i>FTO</i> might have an impact on their levels. Therefore, luciferase RNA was used to generate a standard curve and added to the qRT-PCR assays. <i>GAPDH</i> was used as a reference gene for measuring primary miRNAs transcript levels. Mean ± SD from quadruplicates per assay for three independent cell lines (FTO1C1, FTO2D4 and FTO3C3) are depicted. kd, <i>FTO</i> specific siRNA treated cells, scr, scrambled siRNA treated cells.</p

Topics: Biological Sciences, m 6A, methylation, m 6A demethylase FTO, mirna, rna
Year: 2015
DOI identifier: 10.1371/journal.pone.0118438.g002
OAI identifier: oai:figshare.com:article/1320115
Provided by: FigShare
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