Bx x700

Cultured organoids were dissociated into single cells using TrypLE select (Thermo Fisher Scientific, Waltham, MA, USA) during the third culture passage. Single-cell suspensions were loaded onto the C1 preamp IFC (10–17 μm, Fluidigm San Francisco, CA, USA), for single-cell isolation. Single-cell capture was confirmed under a microscope (BX-X700, Keyence, Osaka, Japan). Pre-amplification of target genes was performed using TaqMan assays (Thermo Fisher Scientific, Waltham, MA, USA), with the IFC controller MX (Fluidigm, San Francisco, CA, USA). Further amplification and data acquirement of the multiplex quantitative PCR analysis was done using the Biomark 48.48. dynamic array IFC and Biomark HD system (Fluidigm, San Francisco, CA, USA), according to manufacturer’s instructions. The set of target genes, and the corresponding gene-specific TaqMan assays analyzed in the study can be found in the Suppl. Table S2.

Alexa647-labeled OVA (Thermo Fisher Scientific, Middletown, VA) with or without SF-10 was administrated intranasally into BALB/c mice as described above. At 15 min after vaccination, the nasal tissues were isolated and cryosection slides were prepared [12 (link), 18 (link)]. The slices were then stained with Hoechst33342 (Doujin Chemical Co., Kumamoto, Japan) and fluorescence images were acquired with a microscope (BX-X700; Keyence Co., Osaka, Japan). At 60 min after vaccination, nasal cells were also isolated by Accutase^®, stained with anti-mouse CD326, CD11c and CD8α Ab (BioLegend), and analyzed by flow cytometry.