Journal: Nature Communications

Article Title: Deciphering bat influenza H18N11 infection dynamics in male Jamaican fruit bats on a single-cell level

doi: 10.1038/s41467-024-48934-6

Figure Lengend Snippet: a Flow cytometry gating strategy for human PBMCs. b Flow cytometry to identify intracellular H18 expression in H18N11-infected myeloid cells (CD11b + ), B cells (CD19 + ), and T cells (CD3 + ) among human PBMC at 24 hpi at an MOI of 5. Representative images from n = 4 independent experiments. c Bar graphs showing quantification of WT H18N11-infected PBMC subsets compared to mock controls. Data are mean ± SD of n = 4 independent experiments. Statistical analysis was performed using a two-tailed Mann–Whitney test ( *p = 0.0286). d Viral growth kinetics in PBMCs infected with WT H18N11 at the indicated MOI. Viral titers were determined by endpoint titration at the indicated time points. Viral titers are the mean ± SD of n = 3 independent experiments. The dashed line indicates the detection limit. e Flow cytometry gating strategy for human monocyte-derived macrophages. f Flow cytometry to detect intracellular H18 expression in monocyte-derived macrophages infected with WT H18N11 or ΔNS1 at an MOI of 5 at 24 hpi. Representative images from n ≥ 5 independent experiments. The bar graph shows the quantification of H18-positive cells. Data are mean ± SD of n = 5 (ΔNS1) or n = 6 (mock, WT H18N11) independent experiments. Statistical analysis was performed using a two-tailed Mann–Whitney test ( **p = 0.0022 for mock vs. WT H18N11 and **p = 0.0043 for mock vs. ΔNS1). g Flow cytometry to determine the viability of monocyte-derived macrophages infected with WT H18N11 or ΔNS1 at an MOI of 5 at 24 hpi. The bar graph shows the quantification of dead cells. Data are mean ± SD of n = 5 (ΔNS1) or n = 6 (mock, WT H18N11) independent experiments. Statistical analysis was performed using a Tukey’s multiple comparison test with single pooled variance ( ***p = 0.0008 for mock vs. WT H18N11, **p = 0.0025 for WT H18N11 vs. ΔNS1). h Subcellular localization of viral NP (green) and H18 (red) antigens in WT H18N11-infected monocyte-derived macrophages was monitored over time by immunofluorescence. Representative images from n = 3 independent experiments. Scale bar, 50 µm. i Viral growth kinetics in monocyte-derived macrophages infected with WT H18N11 or ΔNS1 at the indicated MOI. Viral titers were determined by endpoint titration at the indicated time points. Viral titers are the mean ± SD of n = 4 independent experiments; statistical analysis was performed using Dunnett’s multiple comparison test. * p < 0.05, ** p < 0.01, *** p < 0.001; **** p < 0.0001. Black asterisks, WT H18N11 MOI 5 vs. MOI 0.1; gray asterisks WT H18N11 MOI 5 vs. ΔNS1 MOI 5. The dashed line indicates the detection limit. Source data are provided as a Source Data file.

Article Snippet: Cell surface antigens were stained with BV605-conjugated mouse anti-human CD3 (BioLegend, 1:100), PerCP-Cy5.5-conjugated mouse anti-human CD19 (BioLegend, 1:100), PE/Cy7-conjugated mouse anti-human CD11b (Invitrogen, 1:400) and FITC-conjugated mouse anti-human HLA-DR (BioLegend, 1:100) in FACS buffer.

Techniques: Flow Cytometry, Expressing, Infection, Two Tailed Test, MANN-WHITNEY, Titration, Derivative Assay, Comparison, Immunofluorescence

Journal: Communications Biology

Article Title: Repeated dosing improves oncolytic rhabdovirus therapy in mice via interactions with intravascular monocytes

doi: 10.1038/s42003-022-04254-3

Figure Lengend Snippet: a – h Intravital imaging of CT26 LacZ tumor microenvironment after initial ( a – d ) or second dose ( e – h ) of VSV-AF647. Virus (blue, arrows) delivered as a first dose interacts with tumor cells ( a ), endothelium ( b ), neutrophils ( c ), and other leukocytes ( d ). Virus delivered as a second dose is mainly captured by monocytes defined as CD11b + Ly6G - cells ( e ), expressing Ly6C ( f ), CD169 ( g ), and F4/80 ( h ). Vessels counterstained with FITC-BSA (gray) and delineated by white dashed lines. Scale bar, 25 µm. i Quantification of VSV microdistribution in tumor microenvironment after single or repeated VSV administration. Results are shown as percentage of total VSV-bound cells and plotted as mean ± SEM ( n = 7; two-way ANOVA followed by Sidak’s multiple comparisons test). j FC analysis of intravascular leukocytes in tumor samples at the time of first or second VSV treatment. Intravascular fraction of leukocytes is identified by anti-CD45 injected into the tail vein 10 minutes before tissue collection. Results are shown as percentage of intravascular cells and plotted as mean ± SEM ( n = 4; unpaired t-test). k FC analysis of blood samples collected from untreated mice or 48 h post VSV i.v. injection (10 6 PFU). Results are shown as percentage of CD45 + cells and plotted as mean ± SEM ( n = 4; unpaired t-test). l FC analysis of select leukocyte populations in CT26 LacZ tumor at the time of first or second VSV treatment (see also Supplementary Fig. ). Results are shown as percentage of CD45 + cells and plotted as mean ± SEM (unpaired t -test).

Article Snippet: Rat anti-mouse CD8a eFluor® 660 (53-6.7, 0.2 mg/ml), PE/Cy7-conjugated rat anti-mouse CD11b (M1/70, 0.2 mg/ml) were purchased from eBioscience (San Diego, CA).

Techniques: Imaging, Expressing, Injection

Journal: Journal of Immunology Research

Article Title: M1-Polarized Macrophages Promote Self-Renewing Phenotype of Hepatic Progenitor Cells with Jagged1-Notch Signalling Involved: Relevance in Primary Sclerosing Cholangitis

doi: 10.1155/2018/4807145

Figure Lengend Snippet: Antibodies used for fluorescence studies and Western blot analysis.

Article Snippet: APC-Cy7-conjugated anti- mouse CD11b , Rat mAb , BD Biosciences Catalog No. 557657 , 1 : 100.

Techniques: Fluorescence, Western Blot