Fig 1.
Representative H&E staining of the brains from R.australlis-infected ANXA2-KO and WT mice.
Yellow arrows indicate the presence of focal hemorrhagic lesions. A, mock WT; B&C, mock AXNA2-KO; D-F, R. australis-infected WT; G-L), R. australis-infected AXNA2-KO mice. Scale bar: 20 μm.
Table 1.
Upregulated proteins and their KEGG pathway interpretation from whole-brain lysate.
Fold change is calculated as (KO-WT)/WT.
Table 2.
Downregulated proteins and their KEGG pathway interpretation from whole-brain lysate.
Fold change is calculated as (KO-WT)/WT.
Table 3.
Upregulated proteins and their KEGG pathway interpretation from brain-derived endosomes.
Fold change is calculated as (KO-WT)/WT.
Table 4.
Downregulated proteins and their KEGG pathway interpretation from brain-derived endosomes.
Fold change is calculated as (KO-WT)/WT.
Fig 2.
Representative H&E staining of brain sections from WT (A&B) and ANXA2-KO (C&D) 5 days post-R. australis infection. Perivascular hemorrhage (yellow arrow) can be observed in infected ANXA2-KO group but not infected WT group. TNFα (E) and IFNγ (F) concentrations in serum at 2,4,5 days post-R. australis infection. Relative R. australis DNA copies (G) extracted from the brain of WT and ANXA2-KO mice quantified by rt-qPCR. No significant difference was found. Error bar stands for standard deviation. Scale bar: 20 μm.
Fig 3.
Representative IF staining of SFG rickettsiae (green) in livers, brains, and lungs from WT and ANXA2-KO mice with nuclei of host cells counter-stained with DAPI (blue). The areas indicated by the arrows are enlarged and distinguish rickettsial (green) staining (boxed inserts). Scale bars, 20 μm.
Fig 4.
Functional annotations from GO Term CC (A) and KEGG (B) for whole-brain lysate. The gene lists are composed of differentially-expressed proteins from ANXA2 KO and WT mice challenged with R. australis five days p.i. was analyzed using DAVID. Significantly enriched (p<0.05) functional GO term CC and KEGG pathways are listed. Y-axis indicates the significantly altered functional groups belonging to GO Term CC or KEGG pathways, x-axis indicates the number of the identified proteins in each category.
Fig 5.
Functional annotation from Go Term CC (A) and KEGG (B) for isolated endosomes from the brain. Gene list composed of differentially expressed proteins comparing ANXA2 KO and WT challenged by R. australis five days p.i. was analyzed using DAVID. Functional significantly (p<0.05) enriched GO term CC and KEGG pathways are listed. Y-axis indicates the significantly altered functional groups belonging to GO Term CC or KEGG pathways, and the X-axis indicates the number of the identified proteins in each category.
Fig 6.
Visualization of the GO Term CC CC for whole brain lysate (A) and isolated endosome from the brain (B). Cytoscape BiNGO was used to generate the networks. Each node stands for a GO Term CC category, edges are present to connect functional related nodes. CMH-relevant functional groups (i.e., stress fiber and adherens junction) are enlarged and highlighted from the networks. The color bar indicates the range of p value corresponding to node (ANXA2 KO vs. WT in R. australis infection).
Fig 7.
Cell junction associated protein-protein interactome from whole brain lysate (A) and brain derived isolated endosomes (B). Darker nodes are the source node, representing target genes associated with cell junctions. The gray nodes stands for the interacting genes for our target genes.
Fig 8.
(A) Representative IF staining of ZO-1 (left) and occludin (right) in the brains from R.australis/mock-infected/ WT and ANXA2-KO mice, at day 5 p.i.. Yellow arrows indicate the fragmented structures of ZO-1 and occludin, which were mainly seen in infected ANXA2-KO mice. (B) Representative IF staining for occludin in the livers from R.australis/mock-infected/ WT and ANXA2-KO mice, at day 5 p.i.. (C) Representative IF for VE-cadherin in the brains from R.australis/mock-infected/ WT and ANXA2-KO mice, at day 5 p.i. Yellow arrows indicate the fragmented structures of VE-cadherin. Nuclei of mouse cells were counterstained with DAPI (blue). Scale bar: 20um.
Fig 9.
(A) Representative IF for Ebola virus antigen (Green) in the brains, livers, and lungs from WT or ANXA2-KO mice post-Ebola virus challenge. Yellow arrows represent the presence of Ebola virus antigen. (B-E) H&E staining of the brain from WT(B) and ANXA2-KO (C-E) mice infected by Ebola virus. Yellow arrows indicate the presence of perivascular hemorrhage. (F) Representative IF for ZO-1 (left) and Occludin (right) in the brains from WT or ANXA2-KO mice post-Ebola virus challenge. Yellow arrows indicate the position of ZO-1 or occludin, which represent the tight junction structure. ANXA2-KO but not WT Ebola virus-infected mice exhibited the fragmented tight junction structure in the brain. (G) Representative IF for occludin (red) in the livers from WT or ANXA2-KO mice day 10 post Ebola virus challenge. Yellow arrows show the structure of the pericellular occludin, which was relatively intact in Ebola virus-infected WT mice but dramatically fragmented in the Ebola virus-infected ANXA2-KO mice. Nuclei of mouse cells were counterstained with DAPI (blue). Scale bar: 50 μm (A-E), 20 μm (F and G).