(a) The primary macrophages collected from various deficient genotype mice (No. 1&2 and 11&12 for wild-type as the control, No. 3&4 for TLR-9−/−, No. 5&6 for TLR-4−/−, No. 7&8 for TLR-2−/−, or No. 9&10 for MyD88−/−) were treated with 0.1 µg/ml P. gingivalis LPS (No. 3,5,7,9&11) or 0.1 µg/ml E. coli LPS (No. 4,6,8,10&12) for 2 hrs and washed with PBS and cultured overnight. The cell lysate from each test group was subjected to Western blot with antibodies against mSTAT6B (5278), mLITAF or Actin as the control. The assays were performed in triplicate and a representative experiment is presented. (b) Mouse macrophages were transfected with either mLITAF alone or mSTAT6B alone or cotransfected with both mLITAF and mSTAT6B. Proteins from cytoplasm and nucleus of cells were purified separately and subjected to Western blot analysis using antibodies against mLITAF, mSTAT6B (5287), or Actin (control) as indicated by arrows. The assays were performed in triplicate and a representative experiment is presented. The protein band intensity was analyzed using VersaDoc Imaging System model 4000 MP with Quantity One Quantitation Software version 4.6.3 (Bio-Rad). The band intensity from cytoplasm of untreated cells was used as control and assigned 100%. The other bands were evaluated compared to the control and their intensities with % were attached. (c) Mouse macrophages collected from wild-type mouse (No. 1&2) or from LITAF-deficient genotype mice (No. 3&4) were treated with 0.1 µg/ml E.coli LPS (No. 2&4) or untreated as control (No. 1&3) for 2 hrs and washed with PBS and cultured overnight. Proteins from cytoplasm and nucleus of treated cells were purified separately and subjected to Western blot with the antibodies against mLITAF, mSTAT6B (5287), or Actin as control (indicated by arrows). The assays were performed in triplicate and the representative experiment a shown.