Fig 1: Study flow chart and preservation of lung histopathology in LCOs(A) Study workflow. See also Figure S1 and Tables S1–S3.(B) Bright-field images of LCOs (left column), H&E staining of advanced lung cancer-derived LCOs (middle column), and the primary tumor malignant effusions (right column), including the pericardial effusion, pleural effusion (PE), and ascitic effusion (AE). Scale bars, 100 µm.(C) IHC staining of the adenocarcinoma-derived LCOs as well as the original tissue with classic subtype markers TTF-1 (left column), CK7 (middle column), and Napsin A (right column). Scale bar, 50 µm.(D) IHC staining of CK5/6, P40, and P63 in squamous cell carcinoma. Scale bar for the tissue, 50 µm. Scale bar for the LCO, 20 µm.(E) IHC staining of TTF-1, Syn, CgA, and CD56 markers of small cell lung cancer. Scale bar, 50 µm.
Fig 2: Differentiation pattern of intestinal organoids in decellularized extracellular matrix (dECM-HA-ink II) and Matrigel. (a) Whole-mount immunofluorescence of cultured organoids at Day 7 of Passage 1. Showing villi enterocyte marker CK-20, goblet cell marker mucin-2 (MUC-2), enteroendocrine cell marker chromogranin (CHGA), Paneth cell marker lysozyme (LYZ) and proliferation marker Ki-67. Scale bar 100 µm. (b) Ratio of differentiated marker positive cells to other cells per organoid. Mean ± S.D. (n = 10 organoids). Two-sided t-test. *p < 0.05, ***p < 0.001, and ****p < 0.0001
Fig 3: Corin protein expression in the jejunum and colon. Jejunal and colonic sections from WT mice (male, 10–12 weeks old on normal-salt diet) were stained with H&E (left column). Immunohistochemistry was performed using antibodies against corin (2nd column), ChgA (for enteroendocrine cells) (3rd column), and Muc2 (for goblet cells) (4th column). In the negative control, the primary antibody was replaced by normal IgG (right column). Red (2nd column), green (3rd column), and black (4th column) arrowheads indicate corin-, ChgA-, and Muc2-positive cells, respectively. Scale bars: 20 µm. Representative data were from at least three experiments in each group.
Fig 4: Corin, ANP, and Npra protein expression in the mouse colon. Co-immunofluorescent staining was performed in colonic sections from WT and Corin KO mice (male, 10–12 weeks old on normal-salt diet). (A) Co-staining of corin (red), ANP (purple), and ChgA (green) in enteroendocrine cells of colonic sections from WT (top) and Corin KO (bottom) mice. (B) Co-staining of Npra (red) and E-cad (green) on the luminal surface in colonic sections from WT (top) and Corin KO (bottom) mice. DAPI was used to stain cell nuclei (blue). Scale bars: 20 µm in column 1 and 10 µm in columns 2–5. Representative data were from at least three experiments in each group.
Fig 5: ANP and Npra expression in colon sections from WT and Corin KO mice. Colon samples were from WT and Corin KO mice (male, 10-12 weeks old on normal-salt diet). Immunohistochemical staining was conducted to identify pro-ANP/ANP (ANP) (A) (filled red arrowheads in rows 1–2) and Npra (B) (open red arrowheads in rows 3–4) protein expression in colonic sections form WT (rows 1 and 3) and Corin KO (rows 2 and 4) mice. ChgA (green arrowheads in 3rd column) and Muc2 (black arrowheads in 4th column) staining were included as controls. In the negative control, the primary antibody was replaced by normal IgG. Scale bars: 20 µm. Data are representative of at least three experiments in each group.
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