IL-36a and Pulmonary Inflammation Figure 2. Intratracheal instillation of IL-36a increased the mRNA expression of proinflammatory mediators in the lungs of wild-type C57BL/6 mice. A–H) Transcript expression of early response cytokines (TNFa, IL-1a, IL-1b, IL-36c), the classical IL-1 receptor IL-1R1, the novel IL-1 cytokine cluster receptor IL-36R and the neutrophil specific chemokines CXCL1 and CXCL2 in the lungs of mice 24 h following a single i.t instillation of PBS or 10 mg IL-36a. Transcript expression was evaluated by SYBR-Green based quantitative real-time PCR. I–K) Protein expression of TNFa, IL-1a, IL-1b and CXCL1 in the BAL fluid recovered from mice 24 h following a single i.t instillation of PBS or 10 mg IL-36a. Protein expression was quantified by multiplexed cytometric bead arrays. *Indicates significant differences (P,0.05) compared to PBS treated mice. Data represent mean6SEM from 4– 5 mice/group. doi:10.1371/journal.pone.0045784.g002 PLOS ONE | www.plosone.org 6 September 2012 | Volume 7 | Issue 9 | e45784