Characteristics
subject status: healthy donor
cell type: platelet
gender: Female
treatment: SARS-CoV-2 E protein
Treatment protocol
Platelets were isolated from peripheral blood of healthy donors and treated with SARS-CoV-2 E protein or PBS, respectively.
Extracted molecule
total RNA
Extraction protocol
Fresh blood collected was subjected to platelet isolation within 1 hours after blood collection. Platelets were incubated with SARS-CoV-2 E protein or PBS for 1 hour at 37℃. Then the platelet pellets were isolated by two continuous steps of centrifugation, followed by lysis in QIAzol (QIAGEN) and extracted the total RNA using the miRNeasy Mini Kit (QIAGEN). Total RNA was qualified and quantified using a Nano Drop and Agilent 2100 Bioanalyzer (Thermo Fisher Scientific, MA, USA).
Totol RNA of platelets was isolated by using the miRNeasy Mini Kit (QIAGEN). Platelet RNA was eluated in 30 μL elution buffer.Total RNA was qualified and quantified using a Nano Drop and Agilent 2100 Bioanalyzer (Thermo Fisher Scientific, MA, USA), and included as a quality standard for subsequent experiments only platelet RNA samples with a RIN-value >7 and/or distinctive rRNA curves. The double stranded PCR product from previous step was heat denatured and circularized by the splint oligo sequence. The single strand circle DNA (ssCir DNA) was formatted as the final library. The final library was amplified with phi29(Thermo Fisher Scientific, MA, USA) to make DNA nanoball (DNB) which had more than 300 copies of one molecular, DNBs were loaded into the patterned nanoarray and single end 50 bases reads were generated on BGISEQ500 platform (BGI-Shenzhen, China).
Data processing
RNA sequencing was performed on the BGISEQ500 platform (BGI-Shenzhen, China) and the sequencing data were filtered with SOAPnuke (v1.5.2).
Clean reads were mapped to the reference genome (Species: Homo_sapiens; Source: NCBI; Version: GCF_000001405.39_GRCh38.p13) using HISAT2 (v2.0.4).
Differential expression genes (DEGs) analysis of two groups ( platelets treated with SARS-CoV-2 and platelets treated with PBS) was carried out by DESeq2 (v1.4.5) with log2foldchange >1.0 and Q value ≤0.05. GO (
http://www.geneontology.org/
) enrichment analyses of annotated differentially expressed genes were performed. A P value <0.05 was considered statistical significance.
Assembly: reference genome (Species: Homo_sapiens; Source: NCBI; Version: GCF_000001405.39_GRCh38.p13)
Supplementary files format and content: tab-delimited text files include FPKM values for each Sample
Series (1)
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Supplementary file
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Download
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File type/resource
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GSM6600252_E-3.txt.gz
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259.7 Kb
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(ftp)
(http)
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SRA Run Selector
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Raw data are available in SRA
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Processed data provided as supplementary file
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