Figure 2. Time-dependent TNF--induced COX-2 mRNA and protein in synovial fibroblasts from TMJ. (A) Cells were incubated with 20 ng/mL of TNF- for various time intervals. Total RNA was then isolated from cells and analyzed for COX-2 and GAPDH mRNA by RT-PCR. (B) Cells were incubated with 20 ng/mL of TNF- for various time intervals, and the whole-cell lysates were prepared and subjected to Western blotting with antibody specific for COX-2 and actin. The data represent 1 of 3 separate experiments with similar results.