Fig. 3: mIF-based dissection of immune–epithelial interactions during TCB-mediated toxicity in intestinal organoids.
a, Representative single tiles (×20 magnification) of the 7-plex mIF images across all EpCAM TCB and NT TCB treatments (10 µg ml−1) across 0–72 h time course. PanCK+ organoids are surrounded by CD4+ (orange), CD8+ (turquoise), CD14+ (red) and CD20+ (yellow) immune cells. Caspase-3 (green) captures TCB-mediated off-tumour toxicity in the organoids. Nuclei are stained with DAPI (blue). Scale bar, 100 µm. b, An image at 24 h post EpCAM TCB administration highlights concentric partitioning in 25 μm margins around the initial ROI of the colon organoid (zone 2, red solid line): one inward margin (zone 1), two outward margins (zones 3 and 4). Solid line, inclusion; dotted line, exclusion. Scale bar, 100 µm. c, Mean of the absolute counts of CD4+ and CD8+ T lymphocytes in the zones around individual organoids across time highlights infiltration upon EpCAM TCB (10 µg ml−1) application. All displayed experiments in this figure were replicated at least three times, yielding similar results.
