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E-GEOD-52992 - Rsk2 controls synovial fibroblast hyperplasia and the course of arthritis
Released on 4 December 2014, last updated on 6 December 2014
To understand differences in the pathogenesis of synovial hyperplasia during TNF-induced arthritis, we compared the global gene expression of hTNFtg and hTNFtg;Rsk2-/y primary synovial fibroblasts. Murine fibroblast-like synoviocytes (FLS) were isolated from the hind paws of hTNFtg and hTNFtg;Rsk2-/y (background: C57BL/6) 6-9 week-old mice via collagenase digestion. The cells of two mice were pooled and cultivated for at least 3 passages before usage. For characterization, the cells were tested for the surface expression of CD90.2 and VCAM1 by flow cytometric analysis, whereas CD11b as a monocytic marker had to be absent. Total RNA was extracted using the RNeasy Mini kit (Qiagen). The integrity and amount of isolated RNA was assessed for each sample using an Agilent 2100 Bioanalyzer (Agilent, Waldbronn, Germany) and a NanoDrop ND-1000 spectrophotometer (NanoDrop Technologies, Wilmington, DE). Double-stranded complementary RNA was synthesized from 1 µg total RNA using Message AmpII Biotin (Ambion, USA). Fifteen micrograms of fragmented cRNA of each sample were hybridized to six MG_U430_2 GeneChips (Affymetrix). Hybridization was performed in a Hybridization Oven 640, and chips were washed and stained in the Fluidics Station 450 (both Affymetrix). Finally, the arrays were scanned with a GeneChip Scanner 3000 using the GCOS software, version 1.4, both Affymetrix. All relevant GCOS data of quality checked microarrays were analyzed with High Performance Chip Data Analysis (HPCDA, unpublished), using the BioRetis database (www.bioretis-analysis.de), as described and validated previously.
transcription profiling by array
Joachim R. Grün <Gruen@DRFZ.de>, Andreas Grützkau, Anja Derer, Axel J Hueber, Bettina Grötsch, Bettina Sehnert, Christina Böhm, Georg Schett, Jean-Pierre David, Joachim R Grün, Kirsten Neubert, Michael Stock, Sybille Böhm
SiPaGene: A new repository for instant online retrieval, sharing and meta-analyses of GeneChip expression data. Menssen A, Edinger G, Grï¿½n JR, Haase U, Baumgrass R, Grï¿½tzkau A, Radbruch A, Burmester GR, Hï¿½upl T. , PMID:19265543
Rsk2 controls synovial fibroblast hyperplasia and the course of arthritis. Derer A, Bï¿½hm C, Grï¿½tsch B, Grï¿½n JR, Grï¿½tzkau A, Stock M, Bï¿½hm S, Sehnert B, Gaipl U, Schett G, Hueber AJ, David JP. , PMID:25414238