Transcriptome of Gene Expression
To identify transcription factors (TF) that are important for the maintenance of the odontoblast phenotype, the dental mesenchymal cells (DMC) were isolated from C57BL/6J mouse tooth germ. The DMCs were divided two types, one is collected immediately isolation and the another is collected after 2 days cultures in vitro condition. These cells were comprehensively evaluated and compared using RNA-sequencing (RNA-seq). The data was compared to day0 and day 2 samples with twice. A quality check of RNAs showed over 9.6 RIN value. Library preparation and next generation sequencing (NGS) were done by Macrogen (Macrogen Corp., kyouto, Japan). In briefly, TruSeq stranded mRNA LT Sample prep kit (Illumina, San Diego, CA, USA) was used for library preparation, and NovaSeq6000 system (Illumina) was used for NGS. Data quality was confirmed using Trimmomatic (ver. 0.38,http://www.usadellab. org/cms/?page=trimmomatic). The sequence results were mapped to the reference genomic sequence UCSC mm10 using HISAT2 (ver.2.1.0,https://ccb.jhu. edu/software/ hisat2/index. shtml). StringTie (ver. 1.3.4d, https://ccb.jhu.edu/software/stringtie/) was used to assemble the strings after read mapping. The expression profile was calculated for each sample and transcript/gene as read count and TPM (fragments per kilobase of transcript per million mapped reads).