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. 2023 Mar 15;13:1125901. doi: 10.3389/fcimb.2023.1125901

Figure 5.

Figure 5

Alginate production blocked receptor-ligand binding. (A) To determine if alginate production affects tethering to macrophages, MH-S macrophages were treated with cytochalasin D to inhibit phagocytosis and infected with FRD1, FRD1131 or FRD1131C’ for 1 h. After incubation, macrophages were washed to remove unbound bacteria and lysed to free any bound bacteria. CFU indicates the number of colony-forming units recovered from macrophages. Binding of FRD1 and FRD1131C’ was significantly reduced compared to FRD1131. Data are shown as mean and standard deviation of a representative triplicate experiment. (B, C) To identify cell surface receptors that bind P. aeruginosa FRD, MH-S macrophages were treated with CD11b (Abcam, ab64347) or CD14 (Abcam, ab6083) blocking antibodies and infected with FRD1 (B) or FRD1131 (C) for 1 h. A rat IgG2b isotype control (BD Biosciences, 16061) was used which would presumably function similarly as a control for both the rat (CD11b) and murine (CD14) antibodies. CFU indicates the number of colony-forming units recovered from macrophages. Both antibodies significantly inhibited phagocytosis of FRD1131 compared to the isotype control. However, phagocytosis of FRD1 was not significantly inhibited by either antibody. Data are shown as mean and standard deviation of a representative triplicate experiment. Statistical significance was determined by ANOVA (* P < 0.05, ** P < 0.01, *** P < 0.001).