ena-STUDY-Department of Cardiology, the First Affiliated Hospital of Kunming Medical University-05-08-2021-09:38:49:319-981
Transcriptome sequencing revealed a key signaling pathway in which ECSW improves EPCs function after hypoxia
EPCs were exposed to hypoxic conditions (1% O2, 95% N2, 5% CO2) and starved with 1% fetal bovine serum in EBM-2 medium for 24 hours. Cells incubated under normoxic conditions (95% O2, 5% CO2) and EGM-2 MV medium for 24 hours were used as controls. EPCs were cultured and divided into 3 experimental groups (n=3): (1) control, (2) hypoxia, and (3) hypoxia +ECSW. Paired-end sequencing was performed on an Illumina Hiseq 4000 at LC Sciences, USA, following the vendor's protocol.StringTie was used to analyze mRNA expression levels by calculating FPKM. Differentially expressed mRNAs and genes were selected based on log2 (fold change) >1 or log2 (fold change) <-1 and p value < 0.05).We performed GO enrichment analysis to predict the functions and mechanisms of mRNA and differentially expressed genes. Significant GO terms were defined as those with P-value < 0.05. In addition, KEGG enrichment analysis to predict the signaling pathways in which mRNA may participate was conducted using KEGG Orthology Based Annotation System (KOBAS) v3.0 software. Significant signaling pathways with P-value < 0.05 were included in the analysis.
EPCs were exposed to hypoxic conditions (1% O2, 95% N2, 5% CO2) and starved with 1% fetal bovine serum in EBM-2 medium for 24 hours. Cells incubated under normoxic conditions (95% O2, 5% CO2) and EGM-2 MV medium for 24 hours were used as controls. EPCs were cultured and divided into 3 experimental groups (n=3): (1) control, (2) hypoxia, and (3) hypoxia +ECSW. Paired-end sequencing was performed on an Illumina Hiseq 4000 at LC Sciences, USA, following the vendor's protocol.StringTie was used to analyze mRNA expression levels by calculating FPKM. Differentially expressed mRNAs and genes were selected based on log2 (fold change) >1 or log2 (fold change) <-1 and p value < 0.05).We performed GO enrichment analysis to predict the functions and mechanisms of mRNA and differentially expressed genes. Significant GO terms were defined as those with P-value < 0.05. In addition, KEGG enrichment analysis to predict the signaling pathways in which mRNA may participate was conducted using KEGG Orthology Based Annotation System (KOBAS) v3.0 software. Significant signaling pathways with P-value < 0.05 were included in the analysis.