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. 2022 Feb 22;82(10):1953–1968. doi: 10.1158/0008-5472.CAN-21-0933

Figure 1.

Figure 1. Liver micrometastases of human colorectal cancers (CRC) are devoid of classical SC markers. A, Graphical representation of liver tissue strips extending from macrometastases into adjacent liver tissue. B, Liver tissue strip stained for GPA33 to identify macro- and micrometastases (arrowheads). Dashed lines indicates 1 mm distance to macrometastasis. Scale bar, 5 mm (50 µm in close-ups). C, Distance of identified micrometastases to their respective macrometastasis. Each dot represents one lesion. D, Bar graph summarizing OLFM4 expression patterns in all macro- and micrometastases as either completely absent (green) or heterogeneous (orange). E, Representative IHC stainings of a macro- and micrometastasis of the same patient. Macrometastasis shows heterogeneous OLFM4 expression. Close-ups, orange boxes. Scale bars, 100 µm. F, Experimental setup for spontaneous metastasis formation in mice using orthotopic transplantation of human CRC PDOs with analysis after 9–12 weeks. G, Bar graph depicting the presence of STAR+ CSCs in liver macro- or micrometastases, identified and classified (size) through IHC against human CEA. H, Representative images related to G. STAR minigene is unique to xenotransplanted cancer line. Signal outside CEA-marked metastasis is background. Scale bars, 50 µm.

Liver micrometastases of human colorectal cancers are devoid of classical SC markers. A, Graphical representation of liver tissue strips extending from macrometastases into adjacent liver tissue. B, Liver tissue strip stained for GPA33 to identify macro- and micrometastases (arrowheads). Dashed line, 1 mm distance to macrometastasis. Scale bar, 5 mm (50 µm in close-ups). C, Distance of identified micrometastases to their respective macrometastasis. Each dot represents one lesion. D, Bar graph summarizing OLFM4 expression patterns in all macro- and micrometastases as either completely absent (green) or heterogeneous (orange). E, Representative IHC stainings of a macro- and micrometastasis of the same patient. Macrometastasis shows heterogeneous OLFM4 expression. Close-ups, orange boxes. Scale bars, 100 µm. F, Experimental setup for spontaneous metastasis formation in mice using orthotopic transplantation of human colorectal cancer PDOs with analysis after 9–12 weeks. G, Bar graph depicting the presence of STAR+ CSCs in liver macro- or micrometastases, identified and classified (size) through IHC against human CEA. H, Representative images related to G. STAR minigene is unique to xenotransplanted cancer line. Signal outside CEA-marked metastasis is background. Scale bars, 50 µm. ***, P < 0.001.