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. 2019 Oct 31;13(6):980–991. doi: 10.1016/j.stemcr.2019.10.002

Figure 3.

Figure 3

Characterization of Phagocytosis and Innate Immune Responses of iMACs by H3N2 Viral Infection

(A) Representative micrographs of iMACs engulfing fluorescein isothiocyanate (FITC)-labeled latex beads. Bright-field (BF), GFP, and merged images (1,000× magnification) are shown.

(B and C) Percentage of CD11b+ iMACs containing FITC-labeled latex beads as determined by flow cytometry (n = 3). (B) Opsonized beads were incubated with iMACs as 1:0.2–1:200 ratio (cells:beads). CD11b+ cells were gated and GFP+ cells were presented as dot plot. (C) Percentage of GFP+ cells, mean ± SD, n = 3.

(D and E) Scanning electron micrograph (D) and transmission electron micrograph (E) of iMACs 1 day after H3N2 infection. Scale bars, 1 μm (100 nm enlarged image; left) and 2 μm (200 nm enlarged image; right). Arrows indicate H3N2 viral particles inside cells.

(F) Flow cytometry analysis of superoxide radicals in MitoSOX-stained iMACs 1 day after H3N2 infection. Bar graph represents mean fluorescence intensity (MFI) in vehicle-treated and H3N2-infected cells (n = 3).

(G–I) IFN-γ (G), IFN-β (H), and IL-6 (I) levels in culture medium (CM) of iMACs, 1 day after H3N2 infection.

(J) Relative fold change (RFC) of mRNA levels of immune response genes, after 4 h H3N2 viral infection in iMACs. (A–I) ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001.

(K and L) Type 1 IFN signaling (K) and IFN-γ signaling (L) in H2N3 infected iMACs. Heatmap of differentially expressed genes (p < 0.05) of iMACs on day 1 after H3N2 infection (MOI 5) detected by RNA-seq. Annotated GO terms are indicated.