Figure 4
Markers of LC3-assisted phagocytosis and lysosomal activity. (A–C) Immunofluorescent images showing hCMEC cultures fixed and stained with anti-LC3B antibody following a 1 h incubation with 107 CFU/ml E. coli EV36-RFP alone (A) or 107 PFU/ml phage K1F-GFP alone (B), or a 1 h incubation with 107 CFU/ml E. coli EV36-RFP followed by a 1 h incubation with 104 PFU/ml phage K1F-GFP (C). DAPI stain is shown in blue and anti-LC3B antibody in white. n = 3 in each case. (D,E) Analysis by flow cytometry. (D–F) Graphs showing LysoTracker MFI (+/− SD, n = 3) of hCMEC cultures incubated with 107 CFU/ml E. coli EV36 alone (D) or 107 PFU/ml phage K1F alone (E) over time, or in combination after 1 h incubation (F). (G,H) Graphs showing anti-hLC3B MFI (+/− SD, n = 3) of hCMEC cultures treated with 107 CFU/ml E. coli EV36 (G) or 107 PFU/ml phage K1F (H) over time, or in combination after 1 h incubation (I). Probability values (p-values) are displayed as p ≤ 0.05 (*), p ≤ 0.01 (**), p ≤ 0.001 (***), p ≤ 0.0001 (****).
