Skip to main content
. 2024 Sep 16;20(13):5043–5055. doi: 10.7150/ijbs.97166

Figure 2.

Figure 2

PAD2 inhibitors promoted THP-1 macrophage towards M2 polarization and increased phagocytosis. 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. (A) The expression of classical cytokine genes (Tnfα, Il6, and Il10) and macrophage polarization signature genes (Nos2, Ccl2, and Mrc1) was quantified by qRT-PCR. (B) The concentrations of TNF-α, IL-6 and IL-10 were determined in cell culture supernatant. (C) Protein expression levels of iNOS and Arg-1 were measured using Western blot analysis. (D) Cells were then fixed and stained for iNOS (red), CD206 (green), and DAPI (blue). (E) 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. The phagocytic capacity of cells was measured using pHrodo Red E. coli BioParticles. Data are representative of three independent experiments expressed as means ± SEM. *p < 0.05 vs Control; ***p < 0.001 vs Control; #p < 0.05 vs PA; ##p < 0.01 vs PA; ###p < 0.001 vs PA.