Journal: Life

Article Title: Potential Use of CTCs as Biomarkers in Renal Cancer Patients

doi: 10.3390/life12010089

Figure Lengend Snippet: A schematic presentation of the CellCollector. Straight ( a ) and spiraled ( b ) medical wire covered with anti-EpCAM (blue) and/or anti-MUC1 (black) antibody.

Article Snippet: Incubation with the anti-MUC1 mouse antibody (Cell Signaling) was performed overnight at 4 °C.

Techniques:

Journal: Life

Article Title: Potential Use of CTCs as Biomarkers in Renal Cancer Patients

doi: 10.3390/life12010089

Figure Lengend Snippet: The expression analysis of the renal normal and tumor tissues. ( a ) The transcriptome microarray analysis with a detailed description of the differentially expressed surface proteins; FPKM—Fragments Per Kilobase Million; ( b ) the protein expression of EpCAM, MUC1 and cytokeratin (green) in the clear cell renal tumor (pT3b) and corresponding control tissues. The nuclei were stained with Hoechst (blue).

Article Snippet: Incubation with the anti-MUC1 mouse antibody (Cell Signaling) was performed overnight at 4 °C.

Techniques: Expressing, Microarray, Control, Staining

Journal: Life

Article Title: Potential Use of CTCs as Biomarkers in Renal Cancer Patients

doi: 10.3390/life12010089

Figure Lengend Snippet: The surface markers in the primary ccRCC cell lines. ( a ) The transcriptome microarray analysis with a detailed description of EpCAM and MUC1 in ccRCC31, ccRCC45, ccRCC122 and ccRCC162; FPKM—Fragments Per Kilobase Million, ( b ) the localization of EpCAM (green) and MUC1 (red) proteins on the surface of the ccRCC45 separate cells and merged cells to visualize the colocalization. The images show protein localization. The nuclei were stained with Hoechst (blue).

Article Snippet: Incubation with the anti-MUC1 mouse antibody (Cell Signaling) was performed overnight at 4 °C.

Techniques: Microarray, Staining

Journal: Life

Article Title: Potential Use of CTCs as Biomarkers in Renal Cancer Patients

doi: 10.3390/life12010089

Figure Lengend Snippet: The statistically significant differential expression of EpCAM and MUC1 was determined with a microarray. All analyzed cell lines were compared with RCC45.

Article Snippet: Incubation with the anti-MUC1 mouse antibody (Cell Signaling) was performed overnight at 4 °C.

Techniques: Quantitative Proteomics, Microarray

Journal: Life

Article Title: Potential Use of CTCs as Biomarkers in Renal Cancer Patients

doi: 10.3390/life12010089

Figure Lengend Snippet: The CTCs captured by the spiraled wire. Both anti-EpCAM and anti-EpCAM/MUC1 antibody-covered CellCollector system wires were applied to the blood samples: G3 chromophiles (chRCC) and clear cells (pT3apNxpM1G2) (ccRCC). The attached cells were stained with antibodies against pan cytokeratin CK8, CK18 and CK19 (green) and CD45 (red). The nuclei were stained with Hoechst (blue).

Article Snippet: Incubation with the anti-MUC1 mouse antibody (Cell Signaling) was performed overnight at 4 °C.

Techniques: Staining

Journal: Life

Article Title: Potential Use of CTCs as Biomarkers in Renal Cancer Patients

doi: 10.3390/life12010089

Figure Lengend Snippet: The comparison of the CellCollector wires functionalized with either anti-EpCAM or anti-EpCAM/anti-MUC1 antibodies. ( a ) The number of CTCs captured by anti-EpCAM and anti-EpCAM/anti-MUC1 antibody-covered wires in the matched paired patient samples. Green line—lung metastasis, blue line—other metastases. ( b ) The presentation of CTC detection by both wires with a median of 4 (range 0–15) for anti-EpCAM-coated wires and a median of 6 (range 1–18) for anti-EpCAM/MUC1-coated wires. The mean is displayed as “+” and the whiskers represent the min and max values.

Article Snippet: Incubation with the anti-MUC1 mouse antibody (Cell Signaling) was performed overnight at 4 °C.

Techniques: Comparison

Journal: Inflammatory bowel diseases

Article Title: Aberrant Mucin Expression Profiles Associate With Pediatric Inflammatory Bowel Disease Presentation and Activity.

doi: 10.1093/ibd/izac217

Figure Lengend Snippet: Figure 1. Expression of mucins in the colon of pediatric inflammatory bowel disease (IBD) patients and non-IBD control subjects. A, Relative messenger RNA (mRNA) expression of MUC1, MUC2, MUC3A, MUC3B, MUC4, and MUC13 in inflamed and noninflamed colonic tissue of IBD patients and non-IBD control subjects. Significant differences are indicated by *P < .05; **P < .01; ***P < .001 (1-way analysis of variance or Kruskal-Wallis, Tukey ,and Dunn multiple comparison post hoc test). B, Immunohistochemical analysis of MUC1, MUC2, MUC3, MUC4, and MUC13 expression in colonic biopsies from non-IBD control subjects and IBD patients. Representative images were selected. Scale bars = 20 μm.

Article Snippet: Five-micrometer cross-sections were deparaffinized, rehydrated, and used for immunohistochemical stainings using target specific primary antibodies and visualization with a secondary streptavidin–horseradish peroxidaseconjugated antibody and AEC (3-amino-9-ethylcarbazole) substrate to detect the expression and localization of MUC1 (AF6298; R&D systems; 1:500), MUC2 (NBP1-31,231; Novus Biologicals; 1:2000), MUC3 (NBP2-44,434; Novus Biologicals; 1:100), MUC4 (NBP1-52193; Novus Biologicals; 1:3000), and MUC13 (MABC209; Merck Millipore; 1:1000).

Techniques: Expressing, Control, Comparison, Immunohistochemical staining

Journal: Inflammatory bowel diseases

Article Title: Aberrant Mucin Expression Profiles Associate With Pediatric Inflammatory Bowel Disease Presentation and Activity.

doi: 10.1093/ibd/izac217

Figure Lengend Snippet: Figure 5. Associations of colonic mucin messenger RNA (mRNA) expression with clinical disease parameters in pediatric inflammatory bowel disease (IBD) patients. A, Correlogram of relative mucin mRNA expression and clinical data in pediatric-onset IBD. Spearman rank order correlation values (r) are shown from red (-1.0) to blue (1.0); r values are indicated by color and square size. P values are indicated by black asterisks (*P < .05; **P < .01; ***P < .001). NI indicates mucin mRNA expression in the noninflamed colon; I indicates mucin mRNA expression in the inflamed colon; I_vs_NI indicates log2 fold change of mucin mRNA expression in inflamed colon relative to the noninflamed colon in the same patient. B-F, Correlation of (B) MUC3B mRNA expression with the Pediatric Ulcerative Colitis Activity Index score (NI: n = 7; I: n = 10), (C) MUC3A mRNA expression with the Pediatric Crohn’s Disease Activity Index score (NI: n = 13; I: n = 8), (D) MUC1 mRNA expression with the Mayo score (NI: n = 8; I: n = 12), (E) MUC4 mRNA expression with the Mayo score (NI: n = 8; I: n = 12), and (F) MUC3A mRNA expression with weight loss (NI: n = 24; I: n = 24). r Values are shown and significance is indicated by asterisks (*P < .05; **P < .01) (See online version for color figure).

Article Snippet: Five-micrometer cross-sections were deparaffinized, rehydrated, and used for immunohistochemical stainings using target specific primary antibodies and visualization with a secondary streptavidin–horseradish peroxidaseconjugated antibody and AEC (3-amino-9-ethylcarbazole) substrate to detect the expression and localization of MUC1 (AF6298; R&D systems; 1:500), MUC2 (NBP1-31,231; Novus Biologicals; 1:2000), MUC3 (NBP2-44,434; Novus Biologicals; 1:100), MUC4 (NBP1-52193; Novus Biologicals; 1:3000), and MUC13 (MABC209; Merck Millipore; 1:1000).

Techniques: Expressing, Activity Assay

Journal: Cancer cell international

Article Title: Therapeutic effect of a MUC1-specific monoclonal antibody-drug conjugates against pancreatic cancer model.

doi: 10.1186/s12935-022-02839-w

Figure Lengend Snippet: Fig. 1 HzMUC1 antibody binds to MUC1 on the cell surface of pancreatic cancer cells. A Lysates from SW1990, Mia-PaCa-2, PATU-8988, PANC-1, CFPAC-1, and Capan-2 pancreatic cancer cell lines, and hTERT-HPNE (a normal epithelial pancreatic cell line) were resolved by SDS-PAGE and immunoblotted with the commercially available anti-MUC1 N-terminal (NT) antibody and reprobed with anti-β-actin antibody as loading control. B Cell lysates from Capan-2, CFPAC-1, and SW1990 cells were immunoprecipitated with human IgG or HzMUC1 antibody and immunoblotted with the anti-MUC1-NT antibody or anti-MUC1-CT antibody. C HzMUC1 antibody binds to MUC1 protein on the cell surface. Capan-2, CFPAC-1 and SW1990 cells grown on coverslips were washed with PBS, incubated with HzMUC1 antibody (1 µg/well) on ice, and subsequently with Alexa 488-conjugated secondary antibody, and examined with fluorescent (upper panels) and phase contrast (lower panels) confocal microscopy. Scale bars, 20 μm

Article Snippet: Anti-MUC1 mouse monoclonal antibody to MUC1-N (clone GP1.4; Catalog No. AM32842PU-S at 1:2000) was obtained from OriGene (Rockville, USA).

Techniques: SDS Page, Control, Immunoprecipitation, Incubation, Confocal Microscopy

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