Fig. 6.

CRC Organoids treated with F. nucleatum were more responsive to PD-L1 blockade than those not treated with F. nucleatum. CRC organoids were mixed with T lymphocytes (10⁵/well) and F. nucleatum (10⁸ CFU) and treated with an anti-PD-L1 mAb or an isotype control mAb for one week. a CRC organoids in different groups. Organoid morphology was examined by staining for E-cadherin (red) and with DAPI (blue). Ki-67 expression was detected by IHC (brown staining). Cell apoptosis was detected by TUNEL staining (green). b The proportions of proliferating cells were detected by Ki-67 staining. c The proportions of apoptotic cells were detected by TUNEL staining. One-way ANOVA and Bonferroni’s multiple comparison test. d, e Flow cytometry was used to detect the proportion of CD8⁺ cells in CRC organoids and the proportion of IFN-γ⁺ cells in CD8⁺ cells in different groups. One-way ANOVA and Bonferroni’s multiple comparison test. f CRC organoids were treated with F. nucleatum for different time course. The indicated proteins were detected by Western blotting. One-way ANOVA and Bonferroni’s multiple comparison test. *P < 0.05; **P < 0.01. Fn, F. nucleatum