Expression of total and phosphorylated (phospho) p42/p44 MAPK, SAPK/JNK, p38 MAPK, and NF-κB p65 in lung tissues.

Forty μg of lung tissue homogenate protein from P. aeruginosa-infected and SPA4 peptide- or vehicle-treated mice were separated on SDS-PAGE gel and immunoblotted for phospho NF-κB-p65 (phospho p65), NF-κB-p65 (p65), phospho p38 MAPK (phospho p38), p38 MAPK (p38), phospho p44/p42 MAPK (phospho p44 and phospho p42), total p42/p44 MAPK (total p42/p44), phospho SAPK/JNK (phospho p54 and phospho p46), and SAPK/JNK. The immunoblots were stripped and re-probed with β actin for protein loading. The immunoblots shown are from lung tissue homogenates of three mice in each group, representative of immunoblots performed in separate samples from 12 mice per group in two experiments (A). The densitometric units of immunoreactive bands for NF-κB and MAPK proteins in (A) were normalized with those of β actin. Average densitometric ratios are shown as mean + SEM (B). Ratios of phospho p65/p65, phospho p38/p38, phospho p54/total SAPK/JNK, and phospho p46/total SAPK/JNK are also shown. The p-values were determined by t-test.