Pad2 deficiency and PAD2 specific inhibitors increased autophagy level. (A-B) THP-1 cells were treated with PMA to induce differentiation for 22-24 hours prior to infection with Pseudomonas aeruginosa (PA) at a multiplicity of 100. Cells were then pre-treated with or without 1 µM AFM32a or AFM41a for 24 hours, then treated with or without PA for 1 hour. Protein expression levels of LC3B, Beclin-1, and P62 were measured using immunofluorescence and western blot analysis. (C-D) The Pad2-/- and WT mice were infected with 2.5×106 CFU PA 19660/mouse for 24 hours. Alveolar macrophages (AMs) of WT and Pad2-/- mice were analyzed by immunofluorescence and western blotting to assess the protein levels of autophagy related proteins, LC3, Beclin-1 and P62. Data are representative of three independent experiments expressed as means ± SEM. *p < 0.05 vs Control; #p < 0.05 vs PA.