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. 2021 Dec 13;6:151. doi: 10.1038/s41541-021-00414-4

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© This is a U.S. government work and not under copyright protection in the U.S.; foreign copyright protection may apply 2021

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Fig. 1 — a Vaccination and sample collection schedule. C57BL/6 mice received a prime (week 0)-boost (week 21) vaccination regimen with SpFN formulated with either AH (SpFN + AH) or ALFQ (SpFN + ALFQ). At days 0, 3, 5, 7, and 10, spleen, lymph nodes draining the left quadriceps (dLN), and blood were collected. At week 6 (day 42) post-first vaccination, spleen, mediastinal, and perfused lungs were collected. At week 23 (day 161), perfused lungs were collected. b The t-SNE display of the APC subsets identified by the gating strategy is shown in Supplementary Fig. 1A. c Pie charts comparing the number of recruited APCs measured in the dLNs at days 3 and 5 in response to the two vaccine formulations (n = 5/group/time point) or from naïve mice (n = 4). Twelve APC subsets were identified. Each slice indicates an APC subset; N indicates the average of the total number of APCs quantified in the dLNs. d, e The number of d conventional (cDC1, cDC2) and plasmacytoid DCs (pDCs) and e Medullary cord and medullary sinus macrophage (MCM and MSM) subsets in both vaccine groups at indicated time points. f Activation of cDCs indicated by CD80 and CD86 and g macrophage subset activation indicated by CD80 and CD40 in both vaccine groups at indicated time points. Bars indicate mean + s.d. SSM subcapsular sinus macrophages, MoDC monocyte-derived DC. Experiments were repeated twice and differences between the two groups were analyzed by using a nonparametric Mann–Whitney U-test with p ≤ 0.05 considered as statistically significant.