Journal: Arthritis Research & Therapy
Article Title: High CXCR3 expression in synovial mast cells associated with CXCL9 and CXCL10 expression in inflammatory synovial tissues of patients with rheumatoid arthritis
Figure Lengend Snippet: Analysis of IL-6 mRNA levels within synovial tissue from rheumatoid arthritis (RA) as compared with that from osteoarthritis (OA) patients. Upper panels: quality control of total RNA preparations. Aliquots (300 ng) of total RNA extracted from synovial tissue from RA and OA patients were plotted on a RNA 6000 Nano-LabChip. Quality of RNA was scanned using a 2100 bioanalyzer. RNA gel electropherograms show the presence of 28S and 18S ribosomal units, indicating intact RNA of the investigated samples. Lower panels: differential IL-6 mRNA levels were determined by semiquantitative reverse transcription polymerase chain reaction (PCR). The figure shows a representative analysis of eight cDNA samples derived from patients with RA and of eight cDNA samples from patients with OA. cDNA samples were adjusted to equal glyceraldehyde-3-phosphate dehydrogenase (G3PDH) levels, performed by competitive PCR using an internal standard (see Materials and methods). Numbered lanes correspond to individual patients within Table 1 .
Article Snippet: Quality of all total RNA samples was controlled by a 2100 bioanalyzer according to a RNA 6000 Nano-LabChip Kit procedure (Agilent Technologies, Palo Alto, CA, USA), using 0.3 μg of each total RNA. cDNA was synthesized from 1 μg total RNA in a 20 μl reaction using 200 U Superscript™ II reverse transcriptase (Life Technologies), 500 μmol/l of each deoxynucleotide, 5 mmol/l DTT and 0.5 μg of oligo(dT)15 (Invitek, Berlin, Germany).
Techniques: Reverse Transcription Polymerase Chain Reaction, Polymerase Chain Reaction, Derivative Assay