human mouse cd11b pe cy7  (Multi Sciences (Lianke) Biotech Co Ltd)

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