Inhibition of p38 and p44/42 MAPK signaling by siRNA transfection.

<p>OASF (n = 6) were transfected with siRNA targeting p38 MAPK, p44/42 MAPK and scrambled siRNA (control) before ATP (100 µM) stimulation for 2 h. <b>A</b>. Decreased p38 and p44/42 MAPK expression after siRNA transfection was verified by Western blot analysis. <b>B</b>. Densitometric analysis of Western blot results. <b>C</b>. The ATP-induced BDNF mRNA expression was reduced in p38 MAPK siRNA transfected OASF but not in p44/42 MAPK siRNA transfected cells. Data are shown as means ± standard deviations<b>.</b> *, p<0.05.</p

Purinergic receptor P2X, but not P2Y agonists induce BDNF expression.

<p>OASF (n = 8) and RASF (n = 3) were stimulated for 2 h with 100 µM ATP, ADP and UTP, respectively. BDNF mRNA expression was increased by ATP and ADP, but not by UTP. Data are shown as means ± standard deviations. N.s., not significant; *, p<0.05, ***, p<0.001.</p

Activation of MAPK pathways in OASF after ATP stimulation.

<p><b>A</b>. OASF were stimulated with ATP (100 µM) for 10 minutes, 2 h, 5 h and 24 h. Western blot analysis of p38 and p44/42 MAPK, as well as phosphorylated p-p38 and p-p44/42 MAPK showed kinase activation after 10 minutes ATP stimulation. <b>B</b>. OASF (n = 10) and RASF (n = 3) were treated with the kinase inhibitors FR180204, SB203580 and PD98059, respectively 15 min before ATP (100 µM) stimulation for 2 h. Data are shown as means ± standard deviations<b>.</b> *, p<0.05.</p

Relative DREAM gene expression in peripheral blood mononuclear cells from osteoarthritis (OA) patients and healthy controls

Relative gene expression was normalized to GAPDH (glyceraldehyde-3-phosphate dehydrogenase) and is given as delta CT (dCT) value, with higher values representing lower expression levels. DREAM gene expression was significantly lower in OA patients with a high pain score (visual analog scale [VAS] score of greater than 40; △) compared with healthy controls (○) and with OA patients with a low pain score (VAS score of less than or equal to 40; ∇). No significant differences were observed between healthy controls and OA patients with a VAS score of less than or equal to 40. Statistics: one-way analysis of variance followed by Tukey's honest significant difference (*< 0.05). Ctrl, control; DREAM, downstream regulatory element antagonist modulator.<p><b>Copyright information:</b></p><p>Taken from "DREAM is reduced in synovial fibroblasts of patients with chronic arthritic pain: is it a suitable target for peripheral pain management?"</p><p>http://arthritis-research.com/content/10/3/R60</p><p>Arthritis Research & Therapy 2008;10(3):R60-R60.</p><p>Published online 28 May 2008</p><p>PMCID:PMC2483451.</p><p></p

Qualitative results of reverse transcription-polymerase chain reactions (PCRs) using DREAM primer and DREAM nested primer

DREAM amplicons of 409 base pairs (bp) in size in total RNA derived from cerebellum and spinal cord, which served as positive controls. Amplicons of the expected size after reamplification from total RNA isolated from normal synovial fibroblast-like cells (NSFLCs) and osteoarthritis synovial fibroblast-like cells (OA-SFLCSs). DREAM amplicon of 409 bp and the amplicon resulting from nested PCR, starting from the PCR mix, which did not show any product on the agarose gel. The size of the smaller amplicon corresponds to the expected size of 276 bp. Sequence of the amplicon. Positions of primers are highlighted in bold (DREAM forward and reverse) and bold italics (nested DREAM forward and reverse). DREAM, downstream regulatory element antagonist modulator.<p><b>Copyright information:</b></p><p>Taken from "DREAM is reduced in synovial fibroblasts of patients with chronic arthritic pain: is it a suitable target for peripheral pain management?"</p><p>http://arthritis-research.com/content/10/3/R60</p><p>Arthritis Research & Therapy 2008;10(3):R60-R60.</p><p>Published online 28 May 2008</p><p>PMCID:PMC2483451.</p><p></p