Figure 2
From: Regulation of CD154-induced interleukin-12 production in synovial fluid macrophages
RT-PCR analysis of IL-12p35 and IL-12p40 mRNA expression in synovial fluid (SF) macrophages. Total RNA from SF macrophages (SF M∅), either freshly isolated or incubated with CD154 transfectants or control J558L-cells for 6 or 24 hours. RNA was subjected to reverse transcription, and complementary DNA was amplified by primers specific for IL-12p35, IL-12p40, and constitutively expressed β-actin. cDNA specific for IL-12p35 and IL-12p40 was analyzed using Southern blotting. Reverse transcriptase was omitted from the control reactions. Analysis of SF macrophages from two patients is presented. As a positive control, expression of IL-12p35 and p40 mRNA was analyzed in CD154-stimulated DCs derived from SF macrophages. J558L cells, CD154 transfectants, and aqua were studied as negative controls. RT+ and RT-, incubation with and without reverse transcriptase, respectively.