3d gene human mirna oligo chip version 21

Total RNA was extracted from exosomes and cells using 3D-Gene RNA extraction reagent (Toray Industries, Inc., Tokyo, Japan). The miRNA microarray analysis was performed with a 3D-Gene miRNA Labeling Kit and a 3D-Gene Human miRNA Oligo Chip version 21 (Toray Industries Inc., Tokyo, Japan) to detect 2565 human miRNAs. The chip was scanned with a 3D-Gene Scanner (Toray Industries Inc., Tokyo, Japan) and analyzed with a 3D-Gene Extraction Software (Toray Industries Inc., Tokyo, Japan). The digitalized fluorescence signal provided by this software was used as raw data. All normalized data were globally normalized per microarray such that the median signal intensity was adjusted to 25. From these data, miRNAs that were expressed in HL-60/ADR exosomes at greater than 5-fold higher levels than in HL-60 exosomes were listed.

The standardized method of serum miRNA analysis has been published previously [11 (link)]. Briefly, total RNA was extracted from 300 µL of serum using 3D-Gene^® RNA extraction reagent (Toray Industries, Inc., Tokyo, Japan). A comprehensive miRNA microarray analysis was performed using a 3D-Gene^® Human miRNA Oligo Chip version 21 (Toray Industries, Inc.). The miRNA signal values were normalized to the ratio of the average signal value of three internal control miRNAs (miR-149-3p, miR-2861, and miR-4463) [12 (link)]. The validity of these control miRNAs was confirmed in the present dataset. To identify robust miRNAs, those with a normalized signal value exceeding 64 in more than 50% of the samples in each group were selected. Data sets analyzed in the study were submitted to the National Center for Biotechnology Information Gene Expression Omnibus database under accession number GSE164174.