Fig. 6

CALB2 promotes immunosuppression and metastasis in KPC organoid allograft mice. (A) Schematic diagram depicting KPC organoid xenografts in the C57BL/6 mouse model. KPC organoids were stably transfected with Calb2-OE or control lentivirus and subsequently injected into the spleen of C57BL/6 mice to compare their ability to liver metastasis. (B) IVIS living imaging of the whole body (left) at week 1, 3, and 5, and quantification of the signals at week 5 (n = 5 each group; right). (C) Representative bright-field images and H&E staining images for metastatic lesions (left) and the quantification of metastatic nodules (right). Scale bars: 1 cm (left), 600 μm (middle), 200 μm (right). (D) Metastatic liver sections from C stained with IHC for CALB2 and CXCL14. Scale bars, 500 μm (left), 200 μm (right). (E) Representative microscopy images of bright field and green fluorescence of the indicated KPC organoids following single-cell passage for 5 days. Scale bars, 500 μm. (F) Significantly upregulated pathways (FDR < 0.05 and logFC > 1) identified by DAVID analysis of Calb2-OE liver metastasis (LM) KPC organoids (n = 3) compared to Calb2-OE KPC organoids (n = 3). (G) Volcano plots partially showing differential expression of Calb2, Cxcl14, and EMT-related genes in Calb2-OE-LM KPC organoids compared to Calb2-OE KPC organoids. Red, significant upregulated genes; blue, significantly downregulated genes; grey, non-significantly changed genes. Error bars, mean ± SD; *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001; ns, not significant; by Student’s t test (B and C)