Fig. 6.

Nuclear factor kappa B (NF‐κB) participated in M. pneumoniae lipid‐induced autophagy. (a) RAW264.7 cells were pretreated with 5 mmol/l autophagy inhibitor, 3‐methyladenine (3‐MA), for 1 h and subsequently treated with the lipids (120 μg/ml) for 6 h. Immunofluorescence was performed to detect p65 subunit nuclear translocation of NF‐κB (×100). (b) Data were quantified by counting more than 50 cells in triplicate. (c) RAW264.7 cells were transfected with atg5 and beclin‐1 small interfering RNA (siRNA) for 48 h and then treated with the lipids (120 μg/ml) for 6 h. Immunofluorescence assay was performed to detect NF‐κB nuclear translocation (×100), and data were quantified by counting more than 50 cells in triplicate (d). (e) RAW264.7 cells were pretreated with NF‐κB inhibitor, Bay 11‐7082 (30 μmol/l) for 40 min followed by stimulation with the lipids (120 μg/ml) for 6 h; expression of LC3B and the LC3 puncta was measured by Western blot and immunofluorescence (f,g), respectively. The data shown represent three independent experiments.