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. 2022 Apr 13;13(4):345. doi: 10.1038/s41419-022-04809-6

Fig. 3. Resolving neutrophils trained by 4-PBA exhibit enhanced phagocytosis and bacterial-killing activities.

Fig. 3

A Flow analyses of neutrophil phagocytosis of GFP-labeled E. coli. WT neutrophils programmed with PBS or 4-PBA (1 mM; 24 hours) were subsequently co-incubated with GFP-labeled E. coli for 20 min. Following washing, neutrophils were subjected to flow analyses, and the percentages of GFP-E. coli containing neutrophils were counted and plotted (n = 3). B Analyses of bacterial killing through plating of viable E. coli harvested from lysed neutrophils. WT neutrophils programmed with PBS or 4-PBA (1 mM; 24 hours) were subsequently co-incubated with GFP-labeled E. coli for 30 min. Following washing, neutrophils were lysed and plated on bacterial culture plates. The numbers of viable E. coli were counted (bottom panel) and the CFU (colony-forming units) were plotted (upper panel) (n = 3). The representative image of viable intracellular E. coli from each group was shown. C Analyses of bacterial killing through plating of viable E. coli collected from culture supernatants. WT neutrophils programmed with PBS or 4-PBA (1 mM; 24 hours) were subsequently co-incubated with GFP-labeled E. coli for 30 min. Culture supernatants containing extracellular viable bacteria were plated (bottom panels showing representative culture images) and counted (upper panel) (n = 3). Data were plotted as mean ± SD. *P < 0.05 using two-sided Student’s t test. (A, B, and C).