Figure 2.

TAT-CD137Lct induces IL-6 and TNF production in peritoneal macrophages. (A, B) Peritoneal macrophages (1×10⁵) were treated with indicated concentrations of TAT- EGFP, CD137Lct, TAT-CD137Lct, and TAT-CD137Lct-EGFP and cultured for 4 h. Culture medium was collected and levels of IL-6 (A) and TNF-α (B) were measured using a CBA kit. Heat-inactivated (HI) TAT-CD137Lct was used as a negative control and LPS (10 ng/ml) and CD137-Fc (100 ng/ml) as a positive control. Data are the mean±SD of triplicates. (C) Total RNA was extracted from peritoneal macrophages (1×10⁶) that were treated with 20 ng of protein. Total RNA was subjected to cDNA synthesis. The TNF-α and IL-6 transcripts were amplified using specific primer sets and the PCR products were analyzed using 1.5% agarose gel electrophoresis. GAPDH mRNA expression was used as an internal control. (D) Density of amplified PCR product bands was analyzed using Image J program and normalized against the density of the GAPDH band.