Mirnas serum plasma kit

To further explore the mechanism mediated by macrophage‐derived exosomes, we firstly extracted exosomes from plasma. Then, total RNA was extracted from exosomes using the miRNAs serum/plasma kit (Qiagen, 217,184). The process of sequencing was fully controlled by the data collection software provided by Illumina Xten (Illumina), and the sequence results were collected and analysed by data collection software. Target genes of differentially expressed miRNAs were predicted by the TargetScan 8.0 software. Gene ontology (GO), Kyoto Encyclopedia of genes and genomes (KEGG) enrichment analyses were performed on DAVID 6.8 online databases (https://david.ncifcrf.gov). And the ggplot2 package (version 3.3.5) in R language (version 4.0.5) was adopted to draw bar and bubble plots for visualization, respectively.

The sera obtained were treated with 1/5 of QIAzol, and miRNA isolation was performed using a miRNAs’ Serum/Plasma Kit (Qiagen cat: 217184 Hilden, Germany). The obtained miRNAs were evaluated by spectrophotometric measurements and stored at −20°C. After the isolation, miRNAs were converted to cDNA using an ABM miRNA cDNA Synthesis Kit with poly(A) polymerase tailing (ABM cat: G903 Richmond, Canada).