Skip to main content
. 2020 Nov 11;11:565545. doi: 10.3389/fimmu.2020.565545

Figure 2.

Figure 2

Staphylococcus aureus intracellular survival within PMN is associated with a disruption in autophagic flux. Primary human neutrophils were treated with Bafilomycin A1 (100 nM) or were left untreated and were then infected with pre-opsonized S. aureus PS80 (MoI 10) for 1 h. Following infection, PMN were treated with gentamicin (200 µg/ml) for the times indicated. (A) At each timepoint, PMN lysates were plated onto TSA and CFU enumerated. Data are expressed as Log CFU (n = 4 donors). Statistical analyses were performed using two-way ANOVA with Bonferroni post-tests. *P < 0.05; ***P < 0.001. At each timepoint, PMN protein lysates were probed for LC3 processing and p62 expression using Western immunoblotting (B) and analysed using densitometric analysis (C). Data are expressed as protein expression normalized to β-actin control values for each sample ± SEM (n = 3–4 donors). Black arrows indicate the area of the blot used for densitometry. Statistical analyses were performed using two-way ANOVA with Bonferroni post-tests. **P < 0.01. At 3 h, infected PMN were imaged using transmission electron microscopy (D). Whole, PS80-infected PMN showing phagophore formation indicated by red arrowhead (i) and double-membraned autophagosomes indicated by red arrows (ii) and inset (iii). Original magnification for Ci: 3000x, Cii: 2500x, Ciii: 8000x.