MCL Plays an Anti-Inflammatory Role in Mycobacterium tuberculosis-Induced Immune Response by Inhibiting NF-κB and NLRP3 Inflammasome Activation

Figure 5

MCL inhibits Mtb-triggered NF-κB activation. Raw264.7 cells (1 × 10⁶/well) were seeded in 6-well plates and infected with Mtb for 4 hrs. Cells were washed by PBS and treated with MCL (10 μM) for indicated time courses. Phosphorylation of p65 and β-actin was detected by Western blot (a). The lower bar graph showed the statistical results for phosphorylation levels of NF-κB p65 (a). Data are presented as mean ± SD in at least three independent experiments, . Raw264.7 cells (3 × 10⁵/well) were cotransfected with NF-κB luciferase reporter plasmid and pRL-TKR Renilla luciferase plasmid as previous study [34]. After 30 hr, cells were infected with H37Ra at MOI of 10 and treated with MCL (10 μM) for 12 hr, and the luciferase activities were measured (b). Data are shown with the means ± SD (). . Raw264.7cells (3 × 10⁵/well) were plated in climbing slice and followed by appropriate treatment. Immunofluorescence assay was conducted with rabbit anti-pp65 antibody (green) and DAPI (blue) (c). The results represented three independent experiments.

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