Journal: NPJ precision oncology

Article Title: Chimeric antibody targeting unique epitope on onco-mucin16 reduces tumor burden in pancreatic and lung malignancies.

doi: 10.1038/s41698-023-00423-7

Figure Lengend Snippet: Fig. 4 ch5E6 treatment leads to a substantial decrease in the growth of organoids derived from PC patients or pancreatic and lung cancer genetically engineered mouse models. a PC patient and b KPC mouse organoid staining with ch5E6 indicating MUC16 expression (green) and specific binding of mAb compared to no binding with isotype control mAb huIgG1. The binding of ch5E6 to the ductal cells (yellow) was illustrated by colocalization with CK-19 (red) staining in human and mouse pancreatic tumor organoids. Scale bar 10 µm.The representative images for ch5E6 treated. c Human PC patient. d KPC and (e) KPA mouse-derived organoids obtained by real-time kinetics using Incucyte live cell imaging system. The data were quantitated for organoid counts using essence incucyte software, plotted as a graph of change in organoid counts or area over time (3–5 days) for both ch5E6 and huIgG1 treatments, and is shown in parallel. Error bars indicate SEM. *P < 0.05; **P < 0.01.

Article Snippet: We performed Real time MT-glo assay to determine the effect of anti-MUC16 chimeric mAb5E6 (ch5E6) and isotype control mAb (huIgG1; Bio X Cell).

Techniques: Derivative Assay, Staining, Expressing, Binding Assay, Control, Live Cell Imaging, Software

Journal: NPJ precision oncology

Article Title: Chimeric antibody targeting unique epitope on onco-mucin16 reduces tumor burden in pancreatic and lung malignancies.

doi: 10.1038/s41698-023-00423-7

Figure Lengend Snippet: Fig. 6 Inhibition of EMT by ch5E6 is validated in PC and NSCLC cell line-derived xenografts. a Immunofluorescence analysis showing a decrease in pFAK(Y397) and N-cadherin expression in xenograft tumors of SW1990 cells treated with ch5E6 compared to isotype control mAb huIgG1 group (n = 6–8 fields/tissue: three animals). The data was plotted for changes in fluorescence intensity using GraphPad Prism 9 and is shown in parallel. Nuclei were stained with DAPI. b Immunofluorescence analysis of ch5E6 treated SW1573 cell line-derived xenografts showing a reduction in pFAK(Y397) and N-cadherin levels compared to isotype control mAb huIgG1 group (n = 6–8 fields/tissue: 3 animals). Scale bar, 10 µm; magnified images, 2 µm. No significant changes in the intensity of MUC16 were seen in the ch5E6 treated versus isotype control tumors derived from both cancers. c, d Immunoblot analysis of ch5E6 treated PDAC and NSCLC tumor lysates showing a substantial decrease in phosphorylated levels of FAK(Y397), p70S6K(T389) and N-cadherin as compared to huIgG1 treatment. Error bars indicate SEM. Scale bar, 400 μm; magnified images, 100 μm; *P < 0.05; **P < 0.01.

Article Snippet: We performed Real time MT-glo assay to determine the effect of anti-MUC16 chimeric mAb5E6 (ch5E6) and isotype control mAb (huIgG1; Bio X Cell).

Techniques: Inhibition, Derivative Assay, Expressing, Control, Staining, Western Blot

Journal: NPJ precision oncology

Article Title: Chimeric antibody targeting unique epitope on onco-mucin16 reduces tumor burden in pancreatic and lung malignancies.

doi: 10.1038/s41698-023-00423-7

Figure Lengend Snippet: Fig. 7 MUC16 and N-cadherin are clinically correlated in patient tumors. a, b Representative images and quantitative illustration of immunohistochemical analyses demonstrate a strong positive correlation between MUC16 and N-cadherin (R = 0.84) in both primary PC tumors (n = 10) and liver metastasis (R = 0.99) samples (n = 8). Scale bar, 400 µm; magnified images, 100 µm. *P < 0.05; **P < 0.01. c Representative images of immunofluorescence analysis showing coexpression of MUC16 (green) and N-cadherin (red) in primary PDAC tumors compared to no MUC16 and N-cadherin in normal pancreatic sections. Scale bar, 20 µm; magnified images, 5 µm. d Schematic diagram representing ch5E6 induced downregulation of MUC16 mediated EMT resulting in its anti-tumor potential in PC and NSCLC. Overall, anti- MUC16 chimeric mAb5E6 (ch5E6) binds to the cell surface-tethered domain of MUC16, interferes with oncogenic pFAK/p70S6K/N-cadherin signaling associated with MUC16-mediated EMT, and reduces tumor burden in both PC and NSCLC models.

Article Snippet: We performed Real time MT-glo assay to determine the effect of anti-MUC16 chimeric mAb5E6 (ch5E6) and isotype control mAb (huIgG1; Bio X Cell).

Techniques: Immunohistochemical staining

Journal: Clinics

Article Title: The role of SIGLEC9 in immunosuppression and prognosis in cervical cancer

doi: 10.1016/j.clinsp.2025.100849

Figure Lengend Snippet: PPI map of proteins interacting with SIGLEC9. (A‒D) The PPI network of SIGLEC9 from STRING (A), IntAct (B), BioGIRD (C), Mentha (D). (E) The expression of MUC1 gene in cervical cancer and normal cervical tissue from GEO database. (F) Expression of MUC1 gene in cervical cancer with or without lymph node metastasis from GEO database. (G) The expression of MUC1 gene in cervical cancer and normal cervical tissue from GEPIA database (* p < 0.05).

Article Snippet: The primary antibodies were diluted as follows: rabbit anti-human SIGLEC9 (1:200, Proteintech, China) and MUC1 (1:200, Boster, China) and incubated overnight at 4 °C.

Techniques: Expressing

Journal: Clinics

Article Title: The role of SIGLEC9 in immunosuppression and prognosis in cervical cancer

doi: 10.1016/j.clinsp.2025.100849

Figure Lengend Snippet: Interaction of SIGLEC9 with CD4+ T-cell and MUC1. (A) Immunohistochemical expression of MUC1 in cervical cancer and normal cervical tissues ( n = 40). (B) MUC1 protein expression was analyzed by cell immunofluorescence ( n = 3, three independent experiments). (C) The expression of SIGLEC9 (Green) and CD4 (Red), DAPI (Blue) in normal cervical tissue and in cancer tissues with double immunofluorescence. (D) Correlation between MUC1 (red) and SIGLEC9 (green), DAPI (Blue) in normal tissues ( n = 6) and cancer tissues ( n = 6). (E‒F) Multiplexed immunofluorescence for SIGLEC9 (Red), CD4 (Yellow), and MUC1 (Green), and DAPI (Blue) in cancer tissues and normal tissues. (G‒I) The flow cytometry expression of SIGLEC9+ CD4+ T/CD8+ T-cells and SIGLEC9+M1/M2 in cervical cancer patients ( n = 40) and normal control ( n = 20). ** p < 0.01, *** p < 0.001, and **** p < 0.0001. (J) The cervical cancer patients with a high SIGLEC9+ TAM cell infiltration ( n = 20) had a shorter survival probability than those patients with a low SIGLEC9+ TAM cell infiltration ( n = 20; p = 0.0049).

Article Snippet: The primary antibodies were diluted as follows: rabbit anti-human SIGLEC9 (1:200, Proteintech, China) and MUC1 (1:200, Boster, China) and incubated overnight at 4 °C.

Techniques: Immunohistochemical staining, Expressing, Immunofluorescence, Flow Cytometry, Control

Journal: Frontiers in Medicine

Article Title: Urinary CD8+HLA-DR+ T Cell Abundance Non-invasively Predicts Kidney Transplant Rejection

doi: 10.3389/fmed.2022.928516

Figure Lengend Snippet: Gating strategies for T cell subsets (A) and tubular epithelial cells (TEC) (B) . Isotype controls are displayed as blue, while full stains are represented in red. (C.1) Schematic overview of investigated subsets. Proximal TECs were defined CD10+ and CD13+, while distal TECs were characterized being CD227+ and CD326(EpCAM)+. (C.2) Maturation of naïve T cells into memory T cells. (D) Workflow for epigenetic analysis of urine samples. SSC, side scatter; FSC, forward scatter; TNV, naïve T cells; TEM, T effector memory cells; TCM, T central memory cells; TEMRA, T effector memory cells re-expressing CD45RA.

Article Snippet: The following antibodies were used: for T cells anti-CD3-APCeF780 (eBioscience, SK7, mo IgG1k), -CD4-PEVio770 (Miltenyi Biotec, REA623, REA) -CD8-APC (Biolegend, SK1, mo IgG1k) -CD45RO-PE (Biolegend, UCHL1, mo IgG1k2), -CD45-BUV805 (BD, 3D12, rat IgG1ak), -CCR7-BV421 (Biolegend, G043H7, mo IgG2ak), -HLA-DR-BUV395 (BD, G46-6, mo IgG2ak), -CD28-FITC (Biolegend, CD28.2, mo IgG1k) and for tubular epithelial cells anti-Cytokeratin-FITC (Miltenyi Biotec, CK3-6H5, mo IgG1k), -Vimentin-APC (Miltenyi Biotec, REA409, REA), -CD10-PeVio770 (Miltenyi Biotec, REA877, REA), -CD13-APCVio770 (Miltenyi Biotec, REA263, REA), -CD227-PE (Miltenyi Biotec, REA448, REA), -CD326-BV711 (Biolegend, 9C4, mo IgG2b).

Techniques: Expressing