RNA-Seq of embryonic fibroblasts
The RNA from AF, FF, and EF cells was extracted using RNA-Easy kit (In vitrogen). Libraries for sequencing were prepared using the QIAseq FX Single Cell RNA Library Kit following the manufacturers protocol (Qiagen). Triplicate biological replicates were performed for each group. An Illumina Hiseq 4000 sequencer was used for paired-end sequencing with read length at 150 bp. On average, 25 million reads were obtained for each library. RNA-seq data were aligned to the porcine reference genome (Sscrofa11.1) by using STAR [29]. The identification of differentially expressed genes (DEGs) was performed by using EdgeR [30]. KEGG pathway analysis was performed using clusterProfiler [31].