Fig. 2: Kupffer cells are necessary for the liver anti-metastatic effects of BG.

a Study design for (b). Healthy mice (n = 3) were injected with 5-(4,6-Dichlorotriazinyl) Aminofluorescein (DTAF) labeled β-glucan. Two hours later, mice were euthanized, and livers analyzed. b Frequency of each liver immune subsets bound to β-glucan-DTAF, as measured by FCM. c Frequency of KC and BMDMs bound to β-glucan-DTAF, as measured by FCM in n = 3 healthy mice. d Study design for (e–h). e Representative images of livers from control, BG and BG + DT treated mice 14 days after iPo injection of PDAC-YFP cells. Tissues stained using hematoxylin and eosin (H&E) (top) and mIHC (bottom) to detect CK19 (teal), CD3 (purple), Ki67 (yellow), and nuclei (blue, hematoxylin). Scale bar, 500 μm. Insets show a metastatic lesion. Scale bar, 100μm. f Quantification of PDAC-YFP tumor cells detected in the liver on Day 14 by FCM (n = 8 mice/grp). g PDAC-YFP (CK19⁺) tumor cells detected in liver metastatic lesions on Day 14 by IHC and shown as percent of metastatic lesion area as drawn in (e). (n = 8 mice/group). h Proliferating PDAC-YFP (CK19⁺Ki67⁺) tumor cells detected in liver metastatic lesions on Day 14 by IHC and shown as percent of metastatic lesion area as drawn in (i). Control (n = 138), BG-treated (n = 172) and BG + DT treated (n = 175) individual tumor lesions were analyzed from n = 8 livers per group. Data are representative of 2 independent experiments. Unpaired two-tailed Welch’s t test (c) and one-way ANOVA with Sidak’s test (b, f, g, h). Mean ± SD is shown (b, c, f, g, h). BG-DTAF, β-glucan-5-(4,6-Dichlorotriazinyl) Aminofluorescein; DCs, dendritic cells; KC, Kupffer cells; BMDMs, bone marrow-derived macrophages; iPo, intraportal; DT, diphtheria toxin; BG, β-glucan.