Effect of t10,c12 CLA on microRNA expression.

<p>(A) Total RNA was prepared from three control and three t10,c12 CLA (7 μM) treated cultures of A2780 cells and used to perform small RNA sequencing. After normalization, read counts for known human miRNAs were compared. Mature miRNA transcripts with > 3-fold change (<i>P</i><0.01) are depicted in the heat map. The 46 miRNAs that were differentially expressed are described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0189524#pone.0189524.s003" target="_blank">S1 Fig</a>. (B), shows fold-increase in miR-184 and miR-215 in response to t10,c12 CLA as analyzed by sequencing microarray and qPCR. (C and D) A2780 cells were treated with t10,c12 CLA (7 μM) for 6 h or for the designated time points. The expression of E2F1 was monitored by real-time qPCR (C) and p27/Kip1 was followed by western blotting (D). (E) Intensity of the p27/KIP1 normalized with actin is shown is a graphical representation of the western blots shown in (D). Western blot are representative of three independent experiments.