Journal: Scientific reports

Article Title: Differential expression of CD175 and CA19-9 in pancreatic adenocarcinoma.

doi: 10.1038/s41598-025-86988-8

Figure Lengend Snippet: Fig. 1. Tumor-associated carbohydrate antigens (TACAs) and their detecting antibodies used in this study. Cartoon showing O-GalNAc glycans including the Tn, CA19-9, and CD15s epitopes and their antibody recognition in red dashed line. pp-GalNAc-Ts, polypeptide-N-acetylgalactosaminyl transferases; T-synthase, core 1 β1,3-galactosyl transferase (C1GalT1); Cosmc, C1GalT1-specific molecular chaperone; C2GnTs, Core 2 β1,6-N-acetylglucosaminyl transferases; GTs, glycosyltransferases. ReBaGs6, anti-Tn antibody; CA19-9, anti- CA19-9 antibody; HECA-452, anti-CD15s antibody. Of note, both CA19-9 and CD15s are also expressed on N-glycans.

Article Snippet: After cooling down, sections were washed in water and treated with 0.3% H2O2 (Cat#H1009, Sigma) in deionized water for 10 min, and blocked with 10% goat serum (Cat#PCN5000, Gibco) in TBS + 0.05% Tween-20 (TBST) for 1 h. Sections were stained with a recombinant mouse anti-Tn (anti-CD175) IgM antibody (ReBaGs6, in house40, diluted to 1 μg/ml), mouse anti-SLea (CA19-9) IgG antibody (11C93, Cat#C0075-03E, VWR, diluted to 2 μg/ml), rat anti-SLex (anti-CD15s) IgM antibody (HECA-452, Cat#555,946, BD Bioscience, diluted to 2 μg/ml) or mouse anti-STn (TAG-72, B72.3) IgG antibody (Cat#sc20042, Santa Cruz Biotechnology, diluted to 2 μg/ml) in TBST overnight at 4 °C.

Techniques:

Journal: Scientific reports

Article Title: Differential expression of CD175 and CA19-9 in pancreatic adenocarcinoma.

doi: 10.1038/s41598-025-86988-8

Figure Lengend Snippet: Fig. 2. Comparative expression patterns of TACAs in PDAC. (A) Representative immunohistochemistry (IHC) of benign pancreas, or pancreatic adenocarcinoma (PDAC) stained for Tn, CA19-9, and CD15s antigens. A dark red color indicates the expression of TACAs, and blue represents hematoxylin staining. (B) Comparative H-scores between normal pancreas and PDAC samples for Tn, CA19-9, and CD15s staining (normal, n = 10; PDAC, n = 24, Table 1). P, Kruskal–Wallis one-way analysis of variance (**, p < 0.01). (C) Early pancreatic carcinoma precursor lesions, pancreatic intraepithelial neoplasia (PanIN) and intraductal papillary mucinous neoplasms (IPMN), demonstrate expression of CD175 and CA19-9 (n = 2). All images were captured at 200 × magnification. Scale bars, 100 μm.

Article Snippet: After cooling down, sections were washed in water and treated with 0.3% H2O2 (Cat#H1009, Sigma) in deionized water for 10 min, and blocked with 10% goat serum (Cat#PCN5000, Gibco) in TBS + 0.05% Tween-20 (TBST) for 1 h. Sections were stained with a recombinant mouse anti-Tn (anti-CD175) IgM antibody (ReBaGs6, in house40, diluted to 1 μg/ml), mouse anti-SLea (CA19-9) IgG antibody (11C93, Cat#C0075-03E, VWR, diluted to 2 μg/ml), rat anti-SLex (anti-CD15s) IgM antibody (HECA-452, Cat#555,946, BD Bioscience, diluted to 2 μg/ml) or mouse anti-STn (TAG-72, B72.3) IgG antibody (Cat#sc20042, Santa Cruz Biotechnology, diluted to 2 μg/ml) in TBST overnight at 4 °C.

Techniques: Expressing, Immunohistochemistry, Staining

Journal: Scientific reports

Article Title: Differential expression of CD175 and CA19-9 in pancreatic adenocarcinoma.

doi: 10.1038/s41598-025-86988-8

Figure Lengend Snippet: Fig. 3. Western blots represent unique glycoproteins that carry TACAs in PDAC. Western blots using normal pancreas (24N) and PDAC (39R, 97R, and 103R) tissue extracts with enzymatic treatment with PNGase F (+ PNG), neuraminidase A (+ Neu), or non-treated (NT) analyzed with antibodies to Tn (A), CA19-9 (B), or CD15s (C). Blot images show one representative of two independent experiments with three biological replicates of PDAC (n = 2). See also Supplementary Figure S4 for further information.

Article Snippet: After cooling down, sections were washed in water and treated with 0.3% H2O2 (Cat#H1009, Sigma) in deionized water for 10 min, and blocked with 10% goat serum (Cat#PCN5000, Gibco) in TBS + 0.05% Tween-20 (TBST) for 1 h. Sections were stained with a recombinant mouse anti-Tn (anti-CD175) IgM antibody (ReBaGs6, in house40, diluted to 1 μg/ml), mouse anti-SLea (CA19-9) IgG antibody (11C93, Cat#C0075-03E, VWR, diluted to 2 μg/ml), rat anti-SLex (anti-CD15s) IgM antibody (HECA-452, Cat#555,946, BD Bioscience, diluted to 2 μg/ml) or mouse anti-STn (TAG-72, B72.3) IgG antibody (Cat#sc20042, Santa Cruz Biotechnology, diluted to 2 μg/ml) in TBST overnight at 4 °C.

Techniques: Western Blot

Journal: Scientific reports

Article Title: Differential expression of CD175 and CA19-9 in pancreatic adenocarcinoma.

doi: 10.1038/s41598-025-86988-8

Figure Lengend Snippet: Fig. 4. Spatial differential expression patterns of TACAs in PDAC. Distinct cellular staining of the CD175 (red), CA19-9 (green), and DAPI (blue) in PDAC (109R, A), or normal pancreas (13N, B) sections by confocal microscope. Both Tn + tumor (#1) and CA19-9 + tumor (#2) as indicated in white dashed box from merge of PDAC staining. See Supplementary Figure S5 for other repetitions. Scale bars, 50 μm.

Article Snippet: After cooling down, sections were washed in water and treated with 0.3% H2O2 (Cat#H1009, Sigma) in deionized water for 10 min, and blocked with 10% goat serum (Cat#PCN5000, Gibco) in TBS + 0.05% Tween-20 (TBST) for 1 h. Sections were stained with a recombinant mouse anti-Tn (anti-CD175) IgM antibody (ReBaGs6, in house40, diluted to 1 μg/ml), mouse anti-SLea (CA19-9) IgG antibody (11C93, Cat#C0075-03E, VWR, diluted to 2 μg/ml), rat anti-SLex (anti-CD15s) IgM antibody (HECA-452, Cat#555,946, BD Bioscience, diluted to 2 μg/ml) or mouse anti-STn (TAG-72, B72.3) IgG antibody (Cat#sc20042, Santa Cruz Biotechnology, diluted to 2 μg/ml) in TBST overnight at 4 °C.

Techniques: Quantitative Proteomics, Staining, Microscopy