Human Stem Cell Gene Expression Analysis

A human stem cell RT^{2 (link)} Profiler PCR array (Qiagen Inc., Valencia, CA, USA) was used to evaluate the expression of 84 specific genes. Cells were treated with JNK-IN-8 (2.5 μM) for 48 h, and then total RNA was extracted from the cells using the miRNeasy mini kit (Qiagen Inc.) according to the manufacturer’s instructions. cDNA was synthesized using the RT^{2 (link)} First Strand Kit (Qiagen Inc.) and dispensed into the RT^{2 (link)} Profiler PCR array. Each PCR array included five housekeeping genes (B2M, HPRT1, RPL13A, GAPDH, and ACTB), a control for genomic DNA contamination, a reverse transcription control, and a positive PCR control. The data were analyzed using Qiagen software (http://www.sabiosciences.com/pcr/arrayanalysis.php). The criteria used to define differential expression of genes between each pair of classes tested were P < 0.05 and false discovery rate < 0.2.