Scanner 3000 7g workstation fluidics 450

In order to evaluate the integrity of the RNA, a microfluidic analysis was performed using an Agilent 2100 Bioanalyzer with the RNA6000 nano kit (Agilent Technologies, Palo Alto, CA, USA). For the microarray analysis, we used only RNA samples whose RNA integrity number (RIN) was greater than 8.5. The gene expression was analyzed using a GeneChip® Human Gene 1.0 ST Array (Affymetrix, Santa Clara, CA, USA) containing 764,885 probes (and supporting 28,869 genes). Target cDNA was prepared from 200 ng of total RNA with the Ambion® WT Expression kit (Ambion, Austin, TX, USA) and the Affymetrix® GeneChip® WT Terminal Labeling kit (Affymetrix). Hybridization to the microarrays, washing, staining and scanning were performed using the GeneChip® system (Affymetrix) composed of a Scanner 3000 7G Workstation Fluidics 450 and a Hybridization Oven 645. The scanned image data were processed using the Affymetrix® Expression Console™ Version 1.1. The fold-change for each gene was evaluated using a Gene Expression Analysis with the Partek® Genomics Suite 6.5 software program (Partech, Münster, Germany). Genes with an expression level greater than 2-fold or less than 0.5 were recognized as being significantly different.

The transcriptomes of myometrium, MED12m-positive, and -negative uterine fibroids were analyzed as previously reported^{33 (link),34 (link)}. Total RNAs were isolated from cells by using an RNeasy mini kit (Qiagen). Target cDNA for a microarray was prepared from 250 ng of total RNA with the Ambion WT Expression kit (Ambion, Austin, TX, USA) and the GeneChip WT PLUS reagent kit (Affymetrix). Transcriptomes were analyzed with a GeneChip Human Genome 1.0 ST Array (Affymetrix, Santa Clara, CA, USA) as previously reported^{33 (link),34 (link)}. The microarray was spotted with 21,014 RefSeq genes. Hybridization to the microarrays, washing, staining, and scanning was performed using the GeneChip system (Affymetrix) composed of the Scanner 30,007 G Workstation Fluidics 450 and the Hybridization Oven 645. The scanned image data were processed using a gene expression analysis with the Patrek Genomics Suite 6.5 software program (Partech, Munster, Germany). All expression data were converted to log2 values. Differentially expressed genes (DEGs) were extracted when the expressions in the MED12m-positive or -negative uterine fibroids were higher than 2.0-fold or less than 0.5-fold of that in the myometrium, and p < 0.05 (t-test), and the average expression levels in the tissues with higher expression were more than 100.