Figure 1.

Autophagy-related gene (Atg) 7 interacted with phosphorylated IκBα (p-IκBα) in cells from the murine alveolar macrophage cell line (MH-S). A, Expression levels of Atg7, p-IκBα, and tumor necrosis factor (TNF) α were increased with time after Klebsiella pneumoniae infection (0, 1, 2, or 5 hours) in MH-S and murine alveolar epithelial cell line (MLE-12) cells, as assessed by immunoblotting (IB). Glyceraldehyde 3 phosphate dehydrogenase (GAPDH) was used as a loading control. B, Colocalization of Atg7 and p-IκBα was observed with fluorescence microscopy. MH-S cells were infected with K. pneumoniae at a multiplicity of infection (MOI) of 10:1 for 2 hours. C, Interaction between p-IκBα and Atg7, as detected with a coimmunoprecipitation assay. MH-S cells were infected with K. pneumoniae at an MOI of 10:1 for 0, 2, 5 hours. D, Left, p-IκBα and TNF-α levels were determined with immunoblotting. MH-S cells were transfected with Atg7 small interfering RNA (siRNA) or control (scrambled) siRNA. After 24 hours, cells were infected with K. pneumoniae at an MOI of 10:1 for 1 hour. Right, Nuclear factor κB and TNF-α expression was determined with IB. MH-S cells were transfected with IκBα siRNA or control siRNA. After 24 hours, cells were infected with K. pneumoniae at an MOI of 10:1 for 1 hour. Data were representative of 3 experiments. Abbreviations: +, with; −, without; DIC, differential interference contrast; IgG, immunoglobulin G; IP, immunoprecipitation.