Total RNA including small RNA from sorted cell types was extracted using the mirVana RNA Isolation Kit (Ambion). For whole blood, we used the PAXgene Blood miRNA Kit (Qiagen). For serum samples miRNeasy Serum and Plasma kit (Qiagen) was used and exosomes were processed using the Total Exosome Isolation Reagent and the Total Exosome RNA and Protein Isolation Kit (Life Technologies). All isolation protocols were conducted according to the manufacturers’ instructions, without further modifications.
Extracted total RNAs were combined with a spike-in cocktail (except whole blood samples) as previously described by Hafner et al. (35 (link)). The products were then subjected to Illumina TruSeq Small RNA Sample Preparation protocol to generate small RNA libraries for each sample. Subsequently, the libraries were randomized and pooled with six samples per lane for serum and exosomes, and 24 samples per lane for cell types and whole blood. Sequencing was conducted on an Illumina HiSeq 2500 (1 × 50 bp SR, v3). Raw sequencing reads and quantified read-count data have been deposited at NCBI Gene Expression Omnibus (GEO) (36 (link)) under the accession number GSE100467.
Extracted total RNAs were combined with a spike-in cocktail (except whole blood samples) as previously described by Hafner et al. (35 (link)). The products were then subjected to Illumina TruSeq Small RNA Sample Preparation protocol to generate small RNA libraries for each sample. Subsequently, the libraries were randomized and pooled with six samples per lane for serum and exosomes, and 24 samples per lane for cell types and whole blood. Sequencing was conducted on an Illumina HiSeq 2500 (1 × 50 bp SR, v3). Raw sequencing reads and quantified read-count data have been deposited at NCBI Gene Expression Omnibus (GEO) (36 (link)) under the accession number GSE100467.
