Mir 1246

cDNA from EV-derived small RNAs was synthesised using miRCURY LNA RT kit (Qiagen, Hilden, Germany) following the manufacturer’s specifications. The compatible miRCURY LNA SYBR Green PCR kit was used for real-time qPCR amplification using a CFX Connect Real-Time PCR Detection System (Bio-Rad, Hercules, CA, USA) with the following conditions: 95 °C for 5 min, thereafter 45 amplification cycles at 95 °C for 10 s, and 55 °C for 20 s and 72 °C for 20 s. For all miRNAs, pre-designed miRCURY LNA compatible primers were acquired from Qiagen: miR-1290 (YP02118634), miR-320b (YP02119299), miR-1246 (YP00205630), miR-320a-3p (YP00206042), miR-186-5p (YP00206053), miR-30e-5p (YP00204714), miR-155-5p (YP00204308), miR-99b-5p (YP00205983).

Total RNA from B cells was isolated using TRIzol (Invitrogen Carlsbad, CA, USA). The relative abundance of gene expression was determined by real-time PCR using a Rotor-Gene 3000 (Corbett Research, NSW, Australia). cDNAs were synthesized from 1 μg of total RNA using the miScript Reverse Transcription Kit (Qiagen, Germany). DNA was synthesized from cDNA. Real-time PCR was performed using the Rotor-Gene 3000 Real-time PCR instrument (Corbett Research, Australia). All reactions were run in triplicate. Expression levels of target miRNAs were normalized to RUN6-2 and analyzed with Rotor-Gene Real-Time Analysis Software 6.0. MiR-1246, miR-126, miR-142-3p, miR-142-5p, and RUN6-2 primers were ordered from Qiagen. The information about the primers used for PCR are listed in Additional file 1: Table S2. ΔCt was calculated by subtracting the Ct values for RUN6-2 from the Ct value for the gene of interest. ΔΔCt was calculated by subtracting the control Ct from SLE Ct. The fold change of expression between control and SLE samples was calculated by the equation: 2^−ΔΔCt.