Journal: Journal of Translational Medicine
Article Title: Therapeutic effect of daphnetin on the autoimmune arthritis through demethylation of proapoptotic genes in synovial cells
Figure Lengend Snippet: Effect of daphnetin and 5-aza-dc on expression of DR3, PDCD5, FasL, p53 (A) and DNMT1, DNMT3a, DNMT3b (B) in CIA rat synovial cells. Four experimental groups consisting of untreated cell control, or treated with 5-aza-dc at 20 μM, daphnetin at 40 μg/mL, or combination of 5-aza-dc (20 μM) and daphnetin (40 μg/mL). Cells were cultured, treated and harvested as described in Matreials and Methods. Total RNA was extracted and cDNA was synthesized. After reverse transcription, cDNA was used for real time-PCR. Relative quantification of gene expression was performed by the 2-∆∆Ct method. The results show the mean ± S.D. of six independent experiments. ▲P
Article Snippet: Real-time PCR was performed in the ABI 7500 sequence detection system (Applied Biosystems) with a reaction mixture that consisted of SYBR Green 2 × PCR Master Mix (Tiangen biotech co., LTD., Beijing, China), cDNA template, forward primer and reverse primer.
Techniques: Expressing, Cell Culture, Synthesized, Real-time Polymerase Chain Reaction