Figure 3.

The expression of some 2D HC markers in the invasive front of CRC tissues

(A) A heatmap of Z score-transformed relative expression levels of KRT80, AHNAK2, and AKAP12, normalized by RLE in 9 cultures of 3D MG and 2D HC is shown.

(B) Immunohistochemical images of cytoskeleton-related proteins: KRT80, AHNAK2, and AKAP12 (red) of 3D MG (upper panel) and 2D HC (lower panel) are shown. In MG panels, the region of organoids is indicated with a dashed white line in the left panel. In the right panels, merged images with E-cadherin (gray) are shown. Scale bars, 50 μm.

(C) The protein expression levels of KRT80, AHNAK2, and AKAP12 are verified to be upregulated in 2D HC in each case (#1–3), as indicated by the bar graph. Data are represented as mean ± SD. Statistical significance: ∗p < 0.05, unpaired t-test.

(D) A tiled image of a colorectal cancer surgical specimen is shown. Scale bar, 500 μm. Regions indicated with squares are enlarged in Figure 5E for both the tumoral center and the invasive front.

(E) Differentiated and polarized tumor glands are observed in the tumoral center (top), whereas de-differentiated tumor cells are observed in the invasive front (bottom) after H.E. staining. Immunohistochemical images of KRT80, AHNAK2, and AKAP12 (red) of the tumoral center (upper panel) and the invasive front (lower panel) in the sequential sections are shown. In the H.E. staining map image, the region of the tumoral center is highlighted with a white square, while the invasive front is indicated by a yellow square. In the right panels, merged images with E-cadherin (gray) are shown. Scale bars. 100 μm.

(F) The signal intensity of KRT80, AHNAK2, and AKAP12 in immunofluorescence images are shown by the bar graph in individual cases #1–3. Data are represented as mean ± SD. Statistical significance: ∗p < 0.05, unpaired t-test. See also Figure S2.