Fig 2: Comparative analysis of tumors temporally separated in the same patient. (a) Immunohistochemistry of iPTOs demonstrates robust CD8+ t-cell populations but low granzyme B mediated killing of CK20+ cells. (b) LDH cellular stress analysis determines an increase in pembrolizumab MCC7 iPTO set but not in MCC9, suggesting acquisition of resistance between the two tumors in this patient. All images taken at 40X magnification with scale bar 20 µM. *p < 0.05.

Fig 3: Cancer cells promote apoptosis in hepatocytes in vitro. (a) Schematic illustration of indirect (insert) coculturing system. (b) The left panel shows immunoblotting of cleaved caspase-3 and cleaved PARP-1 in IHH hepatocytes in the presence or absence of indirect contact with colorectal cancer (LS174, SW620 or HT29) cells. The right panel represents the intensity of cleaved caspase-3 and cleaved PARP-1 bands in three independent experiments. (c,d) represent co-immunostaining of IHH hepatocytes indirectly cocultured with HT29 cancer cells using hepatocytes marker, HSA (hepatocyte-specific antigen, green), combined with either cleaved caspase-3 or cleaved PARP-1 (left panel). The right panels represent the quantification of positive pixels. Average pixel intensity was measured from three randomly selected areas for each sample. Results are representative of 3 independent experiments. (e) Schematic showing mixed (direct) coculturing system. (f,g) Representative images of coimmunostaining for IHH hepatocytes cocultured with HT29 cancer cells using a mixing coculturing system. The cells were stained with cancer cell marker, CK20 (cytokeratin 20, green), combined with either cleaved caspase-3 or cleaved PARP-1 (red). The right panels represent the quantification of positive pixels. Average pixel intensity was measured from three randomly selected areas for each sample. Results are representative of 3 independent experiments. The bar chart shows mean values ± SD.