Extended Data Fig. 5: TfR-, CD163-, CD91-bearing EVs derived from S.a-infected mice or BMDM bind iron.
From: Humoral regulation of iron metabolism by extracellular vesicles drives antibacterial response
a-c, Wild-type mice was intraperitoneally injected with S.a and the serum were collected after 24 hours. BMDM were infected with S.a for 24 hours at a MOI of 25 and supernatant was collected. EVs were isolated from the serum or supernatant. a, Western blot analysis of TfR, CD163, CD91 and TSG101 expression in EVs derived from uninfected and infected mouse serum (left), and in EVs released from uninfected or infected BMDM (right). Experiments were repeated three times and representative images are shown. b, Immunoelectron microscopy detection for TfR, CD163, and CD91 antibodies in EVs derived from infected mouse serum (upper) or infected BMDM (bottom). Scale bar, 50 nm. Experiments were repeated twice and representative images are shown. c, Schematic diagram of the overall design of the experiments to test the ability of EVs to bind iron in serum. EVs derived from uninfected mouse serum (Serum EVs group) or S.a-infected mouse serum [Serum (S.a)-EVs group] were added to the serum respectively and then removed by ultracentrifugation. The serum supernatant was collected for further analysis. d, e, Total iron levels (d) and transferrin levels (e) in the serum supernatant after incubation with EVs derived from uninfected or infected mouse serum. n = 3 biologically independent samples. f, Growth of S.a in serum supernatant. n = 5 biologically independent samples. For d-f, data are presented as the mean ± s.d. For d and e, statistical significance was assessed by one-way ANOVA with Tukey’s post-hoc test. For f, statistical significance was assessed by one-way ANOVA with Tukey’s post hoc test and Kruskal-Wallis test.
