The effect of signal inhibitors on human induced pluripotent stem cells in suspension culture conditions

To examine the effect of the effects of PKCbeta and Wnt inhibitors on global gene expression of iPSCs in suspension conditions, RNA-seq experiments were performed. Total RNA was extracted with a FastGene RNA premium kit (Nippon Genetics, Co., Ltd, Tokyo, Japan) and strand-specific library preparation was performed. The prepared library was sequenced by a NovaSeq6000 (Illumina, Inc, CA, USA). Sequencing was performed in a 150 bp x2 paired-end configuration with a data output of about 6 Gb per sample (equivalent to about 20 million paired reads). Library preparation and its sequencing were performed in GENEWIZ (Azenta, MA, USA). The sequencing data were analyzed with a CLC Genomics Workbench (QIAGEN, Hulsterweg, the Neterlands) and the R package edgeR (v3.30.3) to identify differentially regulated genes. Overall design: RNA-seq data from each conditions (n=3)