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. 2022 Dec 22;13:1006998. doi: 10.3389/fimmu.2022.1006998

Figure 4.

Figure 4

Direct contact is required for HLA-DR expression on NK cells after co-culture with IAV-exposed MoDCs. (A) Percentage of HLA-DR+ NK cells and HLA-DR MdFI after 23 h co-culture with mock-treated or IAV-exposed MoDCs at an MOI of 3 as assessed by flow cytometry using an antibody specific to HLA-DR. MoDCs were cultured in direct contact with NK cells (contact), with MoDCs seeded in the dish and NK cells seeded in the transwell with fresh medium (Trans-well) or with conditioned medium (Trans-well (conditioned medium) (n = 3). (B) Schematic of experimental set-up shown in (A). (C) Representative flow histograms of the percentage of the maximum count of HLA-DR on NK cells after exposure to supernatant from mock-treated (top), Cal/09- (middle), or Vic/11- (bottom) exposed MoDCs cultured alone (left panel) or with NK cells (right panel) at a MoDC to NK cell ratio of 4:1 (n = 3). (D) Relative HLA-DR mRNA levels in NK cells re-purified after 23 h co-culture with mock-treated or IAV-exposed MoDCs when compared to NK cell control with RT-qPCR (n = 3). The same shape between conditions indicates that the data point is derived from the same donor. *p < 0.05, ns p > 0.05. Two-way ANOVA, Prism V9.0.0.