FIGURE 1:

CS exposure induced the expression of genes and protein involved in TLR signaling. (A) After C57BL/6J mice (n = 5) were exposed to five cigarettes four times a day for various times (0, 1, 2, 3, and 4 d), the expression levels of the TLR2, TLR4, and TLR9 mRNAs were determined by real-time PCR. Values are expressed as the means ± SD from three independent experiments. *p < 0.05 vs. 0-d treatment. In addition, the expression of the TLR2, TLR4, and TLR9 proteins in the lungs of mice exposed to CS for 3 d was determined by (B) Western and (C) immunofluorescence analyses. The gels were run under the same experimental conditions. (D) Fold increase in the expression of genes involved in TLR signaling in the lungs of mice that had been exposed to CS for 3 d compared with the sham controls using a real-time PCR array. Genes exhibiting at least a fourfold increase in expression in the lungs in response to CS exposure compared with normal samples. (E) The relative expression of the MYD88 mRNA in lungs exposed to CS for 3 d and sham controls was assessed by individual real-time PCR assays to validate the array data. Values are expressed as means ± SD from three independent experiments. *p < 0.05