Fig. 3: β-glucan upregulates antigen presentation pathways in Kupffer cells.

a Study design for (b–i). b UMAP of macrophage populations identified from scRNAseq of control (n = 4) and BG-treated (n = 4) mouse livers. KCs (green) and BMDMs (blue) are defined by Clec4f expression. c, d Volcano plots of differentially expressed genes by pseudobulk analysis among Kupffer cells (c) and BMDMs (d) in control versus BG-treated livers. Annotated upregulated (orange) and downregulated (blue) genes after BG are highlighted. Significance determined by log2FoldChange < −0.5 or > 0.5 and adjusted p-value < 0.05. e Bar graph of selected gene sets identified as enriched by GSEA in KCs after treatment with BG. f Heatmap of IFN-response genes upregulated in KCs after BG. Scale bars indicate log₂[TPM] scaled by row across samples. g Radar plot showing selected gene sets expressed by KCs in control (blue) and BG (orange) treated livers. Axis represents the mean TPM of selected gene set associated genes with range 0 to 172 TPM. h–i Heatmaps of MHC-related (h) and Antigen-presenting cell-related (i) genes upregulated in KCs after BG. Scale bars indicate log₂[TPM] scaled by row across samples. j Quantification of CD206 (left) and MHC class I (H2-Kb/H2-Db, right) expression by BMDMs and KCs in the liver on Day 2 by FCM (n = 5 mice/group). k Quantification of CD206 (left) and CD38 (right) expression by BMDMs and KCs in the liver on Day 14 by FCM (n = 10 mice/group). Two-way ANOVA with correction for multiple comparison (j and k). Mean ± SD is shown (j and k). Flow cytometry data are representative of 2 independent experiments. BMDM, bone marrow derived macrophage; FCM, flow cytometry; iPo, intraportal; KC, Kupffer cell; scRNAseq, single cell RNA sequencing; Ctrl, control; IFN, interferon; MHC, major histocompatibility complex; APC, antigen presenting cell; NES, normalized enrichment score; MFI, mean fluorescence intensity.