Fresh peripheral blood from SLE patients and healthy volunteers was collected and T, B, monocyte, mDC, pDC, naiveB, memoryB and PBMC were isolated at spontaneous state.
Extracted molecule
total RNA
Extraction protocol
Total RNA was extracted using RNAqueous Micro Kit (Thermo Fisher Scientific/Ambion, USA) with RNase-Free DNase Set (Qiagen). RNA Integrity Number (RIN) value was measured using the Agilent Bioanalyzer 2100 (Agilent Technologies, Santa Clara, CA). RNA samples with RIN ≥7.0 were used.
Label
Cy3
Label protocol
Total RNA from a human sample were amplified using the Agilent Low RNA Input Fluorescent Linear Amplification Kit and labeled with Cy3 respectively according to manufacturer’s protocol.
Hybridization protocol
Labeled samples were hybridized to microarray G4110B Human 1A Microarray (V2) for 17hr, 65 deg C.
Scan protocol
Gene expression pixel intensities were extracted by scanning the microarrays with an Agilent Scanner.
Data processing
Data processing and trimming were performed with GeneSpring ver. 12.6.1.
