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Figure 5 | Respiratory Research

Figure 5

From: Mitogen-activated protein kinases and NFκB are involved in SP-A-enhanced responses of macrophages to mycobacteria

Figure 5

SP-A enhances BCG-induced NFκB activation. Panel A: RBMM were obtained from an HIV-1-LTR-luciferase (HLL) transgenic mouse. Mature macrophages were infected for 24 hr with BCG or SP-A-BCG as described in Figure 1. Cells were lysed and luciferase activity was detected by luminometry. Relative light units were corrected by total protein content. *p < 0.05, n = 3. Panel B: RBMM were infected with BCG, SP-A, or SP-A-BCG as described in Figure 1 for 30 min. Nuclear proteins were extracted as described in Methods, and incubated with a 32P-labeled oligonucleotide containing a consensus NFκB binding sequence. Protein-oligonucleotide complexes were then resolved by electrophoresis in a non-reducing polyacrylamide gel. The gel was dried and exposed to film for visualization of bands. LPS at a concentration of 1 μg/ml was run as a positive control (L). Panel C: RBMM were incubated with 1 mM lactacystin for 30 min prior to infection with BCG or SP-A-BCG as described in Figure 1. Nitric oxide was measured in the supernatant after 24 hr. * = p < 0.05, n = 3.

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