Pre amplification kit

10–20 mg of each organ was homogenized in QIAzol Lysis reagent (Qiagen, Germany) using stainless steel beads in TissueLyser II (Qiagen, USA). Total RNA was isolated using Direct-zol RNA MiniPrep kit with on column DNA digestion (Zymo Research Corp., USA). For mRNA analysis, cDNA synthesis was carried out using iScript reverse transcription system kit (BioRad, USA) and quantitative analyses of genes were performed using gene-specific primers on a Bio-Rad iCycler real time machine. Primer sequences for TNFα, MCP1, IL-1β and 18S were same as described28 (link). Cq value was normalized to 18S and fold change was calculated using the delta-delta Ct method. For the detection of miRNAs, TaqMan miRNA Assays (Applied Biosystems) were employed as described previously28 (link). SnoRNA202 (tissue/cells) or synthetic cel-miR-39 (exosomes and plasma) was used to normalize the technical variations between the samples. To enhance miR-155 signal in recipient miR-155 KO mice after the administration of WT plasma (miR-155 enriched) a pre-amplification step was introduced. The cDNA was pre-amplified using pre-amplification kit as described by the supplier (Applied Biosystem, USA) and diluted pre-amplified product was used for real time PCR.

RNA was extracted from human and murine subpopulations, reverse transcribed, and 14 cycles of gene-specific amplification performed using the Applied Biosystem pre-amplification kit with relevant Taqman probe sets (Supplementary Tables 4 and 5). Following amplification the resultant products were loaded in triplicate onto pre-primed 48 × 48 Fluidigm microfluidic dynamic arrays and analysed according to the manufacturer’s instructions. Gene expression was determined relative to GAPDH, HRPT1 or B2m.