Figure 6

Exogenous IL-32α stimulated mRNA expression of IL-6 and MIP-2 independently of NF-κB and MAPK signaling pathways. RAW 264.7 cells were stimulated with rIL-32α in combination with inhibitors for IκB and MAPKs, including DHMEQ, U0126, SB203580, and SP600125, for 6 hours. The amounts of mRNA of TNFα (a), IL-6 (b), and MIP-2 (c) were measured by real-time polymerase chain reaction. rIL-32α alone stimulated RAW 264.7 cells to express IL-6 and MIP-2 as well as TNFα at the mRNA level. None of the inhibitors canceled IL-6 and MIP-2 mRNA expression induced by IL-32α, whereas DHMEQ and U0126 suppressed TNFα mRNA expression. Values are expressed as mean ± standard deviation of triplicate determinations. Ct, control; DHMEQ, dehydroxymethylepoxyquinomicin; GAPDH, glyceraldhyde-3-phosphate dehydrogenase; IκB, inhibitor kappa B; IL, interleukin; LPS, lipopolysaccharide; MAPK, mitogen-activated protein kinase; MIP-2, macrophage inflammatory protein 2; NF-κB, nuclear factor kappa B; n.s., not significant; rIL-32α, recombinant human interleukin-32α protein; TNFα, tumor necrosis factor-alpha.