Quantification of Inflammatory Mediators in Arthritis

The AIA rats were sacrificed on day 23 post-adjuvant induction. The CIA mice were sacrificed on day 33 post-primary immunization. The animals were dissected to obtain synovitis of joint tissues. The total RNA of tissues was extracted in TRNzol Universal reagent by vigorous mechanical shaking. The cDNA synthesis was performed using PrimeScript RT reagent kit and subjected to qPCR reaction using TB Green Premix Ex Taq (Tli RNaseH Plus) kit on ABI 7500 systems (Applied Biosystems, Foster City, CA). The relative mRNA expression was normalized by 2^-ΔΔCt method, and the primer sequences for Tnf, Il1b, Il6, Tab2, Tab3, Ikka, and Actb are listed in Supplementary Table 1.