miR-223-3p Inhibits Antigen Endocytosis and Presentation and Promotes the Tolerogenic Potential of Dendritic Cells through Targeting Mannose Receptor Signaling and Rhob

Figure 5

miR-223-3p regulated DC-initiated CD4+ T cell differentiation Immature mouse DCs were transfected with the miR-223-3p mimic (a, b), mimic control, miR-223-3p inhibitor (c, d), or inhibitor control, at a final concentration of 50 nM. After 24 h, DCs were stimulated with 100 μg/ml OVA for 24 h. Purified CD4+ T cells were cocultured with the transfected DCs at a ratio of 1 : 10 (DC/T cells). After 4 days, cells were collected and stained with anti-CD4-APC, followed by intracellular staining with FITC-conjugated anti-Foxp3, FITC-conjugated anti-IL-17A, PE-conjugated anti-GATA-3, or PE-conjugated anti-T-bet, respectively. The cells were then analyzed by flow cytometry. The bar chart indicated the specific transcription factor of T cells in the gate of CD4+ cells in each group. Data were shown as of three independent experiments. -test, , . (e) The effect of miR-223-3p on CD103 expression in OVA-treated DCs. Immature DCs were transfected with the miR-223-3p inhibitor (50 nM), miR-223-3p mimic (50 nM), or negative control (50 nM) for 24 h, respectively. DCs were incubated with OVA (100 μg/ml) for 24 h, and cells were collected for flow cytometry analysis (gating for CD11c) to determine CD103 expression levels. (f) MFI value presented CD103 expression levels of the three groups. Data were presented as of three independent experiments. Post hoc Tukey’s HSD test, , .

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