The expression of NF-κB and cytokines involved TLR pathway performed by Western blot and proteome profiling assay.

<p>(A) Splenocytes were harvested 6 h after stimulation by PA-MSHA and lysate by RIPA buffer. Expression of NF-κB and phosphor-NF-κB proteins from un-stimulated and PA-MSHA treated splenocytes by Western blot analysis. (B) Spot legend for the cytokine array containing 40 different protein species with duplicates. The array also contains 3 positive control (PC) proteins. For details, see manufacture's protocol (R & D Systems, UK). (C) Negative control for adjuvant stimulation, using medium alone. (D) PA-MSHA (10⁷ CFU/ml) treatment. All arrays were processed under the same conditions after stimulation. (E) Densitometry represents mean corrected for background. Expression levels elevated by more than 1.5-fold relative to control were identified as up-regulated.