Characterisation of fibroblast-like synoviocytes from a murine model of joint inflammation
1 Bone Research Program, ANZAC Research Institute, University of Sydney, Hospital Road, Concord, Sydney, 2139, Australia
2 Centre for Endocrinology, Diabetes and Metabolism, Institute of Biomedical Research, University of Birmingham, Vincent Drive, Birmingham, B15 2TT, UK
3 Department of Immunity and Infection, Institute of Biomedical Research, University of Birmingham, Vincent Drive, Birmingham, B15 2TT, UK
4 Department of Endocrinology & Metabolism, Concord Repatriation Hospital, University of Sydney, Hospital Road, Concord, Sydney, 2139, Australia
Arthritis Research & Therapy 2013, 15:R24 doi:10.1186/ar4158Published: 29 January 2013
Additional file 1:
Unedited mRNA gel images. Unedited gels for the mRNA expression of 18S, prolyl 4-hydroxylase (P4H), CD248, osteocalcin (OCN) and CD68 determined by standard RT-PCR at 35 cycles in one non-inflamed control FLS (wild-type), three K/BxN FLS lines (FLS 1 to 3), primary calvarial osteoblasts (OBs) and the macrophage cell line RAW 264.7 (MΦ).
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Additional file 2:
Positive control images for CD31 and CD68 staining. Expression of CD31 (blue) in K/BxN fibroblast-like synoviocytes (FLS) culture determined by confocal immunofluorescence and in murine lung by immunohistocemistry, CD68 (red) in K/BxN FLS and in RAW CD68 +ve cells determined by confocal immunofluorescence. Nuclei are counterstained in green. Images shown are representative of three separate FLS cell lines.
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Additional file 3:
mRNA expression of CD68, cadherin 11 and IL-6. (a) Fold change in mRNA expression of the macrophage surface marker CD68 in K/BxN fibroblast-like synoviocytes (FLS) at passages 3 and 4 determined by real-time RT-PCR. For each gene product data were normalized for levels of the housekeeping gene 18S rRNA and are presented as fold change in expression (± standard error) relative to the P2 control. Results shown are the combined duplicates of two K/BxN FLS. (b) mRNA ΔCt values for cadherin 11 in three separate wild-type non-inflamed and three K/BxN FLS cell lines after 16 hr treatments with either vehicle or TNF-α at 10 ng/ml. (c) Fold change in IL-6 mRNA expression over 0, 2, 4, 8, 16 and 24 hr following treatment with either corticosterone (100 nmol/l) or TNF-α (10 ng/ml). Results shown are the combined duplicates of three separate FLS lines. Data were normalized for levels of the housekeeping gene 18S rRNA and are presented as fold change in expression (± standard error) relative to the 0 hr control. *P < 0.05, **P < 0.001 versus respective untreated control.
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