Fig. 4

Expression of costimulatory molecules in activated DCs from CLNs. Mice were immunized with PBS, eggshell-derived HAs, MVs or MVs plus eggshell-derived HAs. One week after immunization, the CLNs were isolated, and mononuclear cells were prepared and subjected to flow cytometry. Representative flow cytometry plots (upper panels) and the frequencies of costimulatory molecules (CD40, CD80, and CD86)- or MHC class II (I-A^d)-expressing CD83⁺ cells within the CD11c⁺ DCs (lower panel) are presented. Bar graph data are presented as relative frequencies normalized to those of the PBS-immunized control group. The gating strategy is detailed in Supplementary Fig. 4. A significant difference (exact p-values) between experimental groups and the PBS-treated control group was determined by the unpaired nonparametric Mann‒Whitney U test. EHA; eggshell-derived HAs.