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. 2025 Dec 19;17:761. doi: 10.1038/s41467-025-67437-6

Fig. 1. Glycerol metabolites from L. reuteri inhibit CRC growth in vitro and in vivo, and increase immune cells in colon tumors.

Fig. 1

a Schematic illustration of the culture of L. reuteri in glycerol, the centrifugal collection of glycerol metabolites, and the antitumor activity assessment in different cancer models. b Cell viabilities of NCM460, HUVEC, L-02, HCT116, and MC38 cells after a 24-hour culture with the collected metabolites at different reuterin concentrations (n = 5). c Colony-forming assay of HCT116 and MC38 cells treated with the metabolites at different reuterin concentrations. d Viabilities of MC38 cells after treatment with mixed metabolites (containing 6.25 μg/ml of reuterin) or pretreatment with 1.5 mg/ml NAC for 24 h (n = 5). e The representative microscopic and live/dead staining images of the PDOs derived from MSI-H and MSS CRC patients treated with pure reuterin (1.5 μg/ml) or mixed metabolites (containing 1.5 μg/ml of reuterin) for 3 days. Scale bar, 50 μm. f The organoid diameter after a 3-day treatment with the given agents (n = 10). g Tumor growth curves of subcutaneous MC38 cancer-bearing mice received an intratumoral injection (i.t.) or oral gavage (o.g.) of the given agents (n = 7). h, i The representative immunofluorescence staining images and the corresponding quantification of macrophages (F4/80+), DCs (CD11c+), and CD8+ T cells within the tumor tissues from the mice receiving the given treatments (n = 15 fields from 3 mice per group). Scale bar, 50 μm. The area within the white box is shown magnified. The “n” represents the number of biologically independent samples. Data are shown as mean ± SD; p-value (compared to Ctrl in f; compared to PBS group in g); one-way ANOVA (one-tailed), Tukey’s multiple comparisons test. Source data are provided as a Source Data file.