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

Journal: Investigative Ophthalmology & Visual Science

Article Title: Transmembrane Mucin 1 Blocks Fluorescein Ingress to Corneal Epithelium

doi: 10.1167/iovs.63.2.31

Figure Lengend Snippet: Fluorescein punctate staining are the fluorescein-incorporated corneal epithelial cells of the superficial layer featured with decreased MUC1 expression. (A) The clinical or slit lamp view under cobalt blue light of the eyes of sham-operated rabbit model (lower panel) or dry eye rabbit model ( upper panel ) following fluorescein staining. (B) The fluorescein-stained corneal epithelia prepared as the frozen tissue sections were stained with MUC1 for immunohistochemistry ( brown , left panel ) and immunofluorescence ( middle and right panel ) analyses. For immunofluorescence, dual channels ( red and blue , middle panel ) or triple channels ( red , blue , and green , right panel ) were used. The cell nuclei were counterstained with hematoxylin ( purple , immunohistochemistry) or DAPI ( blue , immunofluorescence). (C) The frozen corneal epithelia section prepared from sham-operated and lacrimal gland-removed rabbits were immunostained with ZO-1 ( red ), nuclear counterstained with DAPI ( blue ), and the fluorescein ingress ( green ) were analyzed through a confocal microscopy. ( red and blue dual channels, middle panel ; red , blue , and green triple channels, right panel ).

Article Snippet: The cryosections were then stained with a biotin-conjugated anti-rabbit MUC1 rabbit antibody (no. NBP1-60046B; Novus Biologicals, Littleton, CO, USA), anti-ZO-1 (no. 33-9100; ThermoFisher Scientific, Waltham, MA, USA), or anti-galectin-3 (GAL3) mouse antibody (no. A3A12, Novus Biologicals) for one hour followed by incubation in a Texas Red conjugate of NeutrAvidin biotin-binding protein (no. A2665; Invitrogen, Carlsbad, CA, USA), Alexa Fluor 594 goat anti-rabbit IgG (H+L) cross-adsorbed secondary antibody (no. A-11012; Invitrogen), or Alexa Fluor 564-conjugated goat anti-mouse IgG antibody (no. A-11003) for one hour in the darkness.

Techniques: Staining, Expressing, Immunohistochemistry, Immunofluorescence, Confocal Microscopy

Journal: Investigative Ophthalmology & Visual Science

Article Title: Transmembrane Mucin 1 Blocks Fluorescein Ingress to Corneal Epithelium

doi: 10.1167/iovs.63.2.31

Figure Lengend Snippet: The fluorescein intensity is reversely correlated with the level of MUC1 in HCECs cultured in MPM. (A) The HCECs that have been grown in KSFM were cultured in MPM for two or five days, and the levels of MUC1, MUC4, and MUC16 mRNA were examined. (B) Representative flow cytometric analyses of MUC1 expressions on cell membranes of HCECs cultured in KSFM ( left ) or MPM ( right ) were presented in contour plots. (C) Representative flow cytometric analyses of fluorescence intensity and MUC1 expression in HCECs cultured in KSFM ( left ) or MPM ( right ) followed by fluorescein staining were presented in contour plots. (D) The HCECs cultured in KSFM or MPM were stained with fluorescein, and the levels of fluorescence ( green ) and MUC1 ( red ) were examined using an immunofluorescence microscopy. Nuclei were stained with DAPI ( blue ). The images were obtained via a confocal microscopy.

Article Snippet: The cryosections were then stained with a biotin-conjugated anti-rabbit MUC1 rabbit antibody (no. NBP1-60046B; Novus Biologicals, Littleton, CO, USA), anti-ZO-1 (no. 33-9100; ThermoFisher Scientific, Waltham, MA, USA), or anti-galectin-3 (GAL3) mouse antibody (no. A3A12, Novus Biologicals) for one hour followed by incubation in a Texas Red conjugate of NeutrAvidin biotin-binding protein (no. A2665; Invitrogen, Carlsbad, CA, USA), Alexa Fluor 594 goat anti-rabbit IgG (H+L) cross-adsorbed secondary antibody (no. A-11012; Invitrogen), or Alexa Fluor 564-conjugated goat anti-mouse IgG antibody (no. A-11003) for one hour in the darkness.

Techniques: Cell Culture, Fluorescence, Expressing, Staining, Immunofluorescence, Microscopy, Confocal Microscopy

Journal: Investigative Ophthalmology & Visual Science

Article Title: Transmembrane Mucin 1 Blocks Fluorescein Ingress to Corneal Epithelium

doi: 10.1167/iovs.63.2.31

Figure Lengend Snippet: Diquafosol treatments preferentially promotes the expression of MUC1 on cell membranes. (A) The HCECs were grown in a culture media containing 0, 0.5, or 1.0 mM diquafosol for four days, and the levels of MUC1, MUC4, and MUC16 mRNA of treated cells were examined using qRT-PCR. (B) Representative flow cytometric analyses of MUC1, MUC4, and MUC16 expression on cell membranes of HCECs treated with 1 mM diquafosol ( red ) or control ( gray ) were presented in overlaid histograms of signal intensity. (C) Representative flow cytometric analyses of fluorescence intensity and MUC1 expression in HCECs treated with sham ( left ) or 1 mM diquafosol ( right ) followed by fluorescein staining were presented in contour plots.

Article Snippet: The cryosections were then stained with a biotin-conjugated anti-rabbit MUC1 rabbit antibody (no. NBP1-60046B; Novus Biologicals, Littleton, CO, USA), anti-ZO-1 (no. 33-9100; ThermoFisher Scientific, Waltham, MA, USA), or anti-galectin-3 (GAL3) mouse antibody (no. A3A12, Novus Biologicals) for one hour followed by incubation in a Texas Red conjugate of NeutrAvidin biotin-binding protein (no. A2665; Invitrogen, Carlsbad, CA, USA), Alexa Fluor 594 goat anti-rabbit IgG (H+L) cross-adsorbed secondary antibody (no. A-11012; Invitrogen), or Alexa Fluor 564-conjugated goat anti-mouse IgG antibody (no. A-11003) for one hour in the darkness.

Techniques: Expressing, Quantitative RT-PCR, Control, Fluorescence, Staining

Journal: Investigative Ophthalmology & Visual Science

Article Title: Transmembrane Mucin 1 Blocks Fluorescein Ingress to Corneal Epithelium

doi: 10.1167/iovs.63.2.31

Figure Lengend Snippet: Knockdown of MUC1 leads to an increase in fluorescein ingress. (A) The mRNA levels of MUC1, MUC4, and MUC16 in HCECs separately transduced with two lentiviral-based shRNA vectors (shMUC1-1 or shMUC1-2) were examined using RT-PCR. The β-actin gene was used as an internal control. (B) Representative flow cytometric analyses of fluorescence intensity and MUC1 expressions in transduced HCECs followed by fluorescein staining were shown in contour plots.

Article Snippet: The cryosections were then stained with a biotin-conjugated anti-rabbit MUC1 rabbit antibody (no. NBP1-60046B; Novus Biologicals, Littleton, CO, USA), anti-ZO-1 (no. 33-9100; ThermoFisher Scientific, Waltham, MA, USA), or anti-galectin-3 (GAL3) mouse antibody (no. A3A12, Novus Biologicals) for one hour followed by incubation in a Texas Red conjugate of NeutrAvidin biotin-binding protein (no. A2665; Invitrogen, Carlsbad, CA, USA), Alexa Fluor 594 goat anti-rabbit IgG (H+L) cross-adsorbed secondary antibody (no. A-11012; Invitrogen), or Alexa Fluor 564-conjugated goat anti-mouse IgG antibody (no. A-11003) for one hour in the darkness.

Techniques: Knockdown, Transduction, shRNA, Reverse Transcription Polymerase Chain Reaction, Control, Fluorescence, Staining

Journal: Investigative Ophthalmology & Visual Science

Article Title: Transmembrane Mucin 1 Blocks Fluorescein Ingress to Corneal Epithelium

doi: 10.1167/iovs.63.2.31

Figure Lengend Snippet: The overexpression of MUC1 lacking the extracellular domain is unable to block fluorescein ingress. (A) Flow cytometric analyses of keratin 3, keratin 12, and vimentin expressions in HCECs cultured in KSFM ( gray ) or MPM ( red ) were presented in overlaid histograms of signal intensity. (B) Schematic diagrams of the wild-type and N-terminally truncated MUC1. (C) Representative flow cytometric analyses of fluorescence intensity and MUC1-C levels in HCECs transfected with the control vector ( left ) or the vector encoded with N-terminally truncated MUC1s ( right ) followed by fluorescein staining were shown in contour plots.

Article Snippet: The cryosections were then stained with a biotin-conjugated anti-rabbit MUC1 rabbit antibody (no. NBP1-60046B; Novus Biologicals, Littleton, CO, USA), anti-ZO-1 (no. 33-9100; ThermoFisher Scientific, Waltham, MA, USA), or anti-galectin-3 (GAL3) mouse antibody (no. A3A12, Novus Biologicals) for one hour followed by incubation in a Texas Red conjugate of NeutrAvidin biotin-binding protein (no. A2665; Invitrogen, Carlsbad, CA, USA), Alexa Fluor 594 goat anti-rabbit IgG (H+L) cross-adsorbed secondary antibody (no. A-11012; Invitrogen), or Alexa Fluor 564-conjugated goat anti-mouse IgG antibody (no. A-11003) for one hour in the darkness.

Techniques: Over Expression, Blocking Assay, Cell Culture, Fluorescence, Transfection, Control, Plasmid Preparation, Staining

Journal: The Journal of clinical endocrinology and metabolism

Article Title: MUC1 as a discriminator between endometrium from fertile and infertile patients with PCOS and endometriosis.

doi: 10.1210/jc.2010-0603

Figure Lengend Snippet: FIG. 1. Expression of MUC1 subunits in secretory phase endometrium. Secretory phase endometrium from fertile (n 33), endometriosis (Endom) (n 25), and ovPCOS (n 26) patients was analyzed for the expression of MUC1ND and MUC1CD subunits as described in Patients and Methods. In parallel, endometrial samples obtained from 15 anovPCOS patients were processed as before to analyze the expression of MUC1 subunits. A, Immunohistochemistry figures. In ovPCOS women, MUC1ND and MUC1CD staining was stronger than in fertile and anovPCOS patients. In anovPCOS women, MUC1ND and MUC1CD staining was significantly reduced in the glandular epithelium compared with fertile patients. Endometriosis patients showed a significantly reduced expression of MUC1CD staining in glandular and luminal epithelium compared with fertile patients. Magnification, 20. Statistical analysis of the immunohistochemistry scores is showed in Fig. 2. B and C, Cumulative scores for each antibody and groups are plotted. Values are expressed as median (R) and interquartile range (box and whisker). Statistical analysis was performed using the Kruskal-Wallis test followed by a Mann-Whitney test. *, P 0.05; **, P 0.01 are considered significant. F, Fertile patients, white box, n 33; anovPCOS patients, gray box, n 15; ovPCOS patients, dark gray box, n 26; E, endometriosis patients, hatched box, n 25.

Article Snippet: A rabbit MUC1CD monoclonal antibody (clone G22-L; Acris Antibodies GmbH, Herford, Germany) was used for immunohistochemistry.

Techniques: Expressing, Immunohistochemistry, Staining, Whisker Assay, MANN-WHITNEY

Journal: The Journal of clinical endocrinology and metabolism

Article Title: MUC1 as a discriminator between endometrium from fertile and infertile patients with PCOS and endometriosis.

doi: 10.1210/jc.2010-0603

Figure Lengend Snippet: FIG. 2. Expression of MUC1 subunits in epithelial cells isolated from endometrial biopsies of fertile and infertile patients. A and B, Protein extracts of confluent epithelial cell monolayers grown from fertile and infertile endometrium were analyzed for the expression of MUC1 subunits as described in Patients and Methods. Protein levels were normalized to GAPDH protein levels. Representative blots are shown. C and D, Densitometric analysis of the blots for both MUC1 subunits was performed using the Quantity One software (Bio-Rad). Values are band density normalized to GAPDH and are expressed as intensity per square millimeter (INT/mm2). Values are expressed as median (R) and interquartile range (box and whisker). Fertile (white box), anovPCOS (gray box), ovPCOS (dark gray box), endometriosis (Endom, hatched box). Statistical analysis was performed using the Kruskal-Wallis test to determine significance between groups, followed by a Mann-Whitney test to compare specific group pairs. In ovPCOS patients (n 6), MUC1 subunit protein expression levels were higher than in fertile (n 6) and anovPCOS (n 6) patients. MUC1ND subunit was found to be expressed significantly higher in fertile patients than in endometriosis patients (n 6). MUC1CD subunit levels were significantly higher in fertile than in anovPCOS and endometriosis patients. E, RNA samples obtained from epithelial cells grown from fertile and infertile endometrium were analyzed for the expression of MUC1 transcript levels as described in Patients and Methods. RNA transcript levels correlated to the protein levels for each group. Values are expressed as average and SD. Statistical analysis of the data was performed using an ANOVA followed by an unpaired t test. *, P 0.05; and **, P 0.01 are considered significant.

Article Snippet: A rabbit MUC1CD monoclonal antibody (clone G22-L; Acris Antibodies GmbH, Herford, Germany) was used for immunohistochemistry.

Techniques: Expressing, Isolation, Software, Whisker Assay, MANN-WHITNEY

Journal: The Journal of Biological Chemistry

Article Title: The protease cathepsin K can debulk the cancer glycocalyx

doi: 10.1016/j.jbc.2026.111206

Figure Lengend Snippet: Only cathepsin K degrades cell-surface mucins on K562s. A, schematic describing the flow cytometry assay used to evaluate the degradation of cell-surface mucins on K562 cells by cathepsins. B, cell viability of cells following enzymatic treatment. Normalized staining for ( C ) MUC1, ( D ) CD43, and ( E ) total mucins via StcE E447D following enzymatic treatments for 3 h at pH 6 in Hanks' balanced salt solution (HBSS). Staining was normalized such that PBS-treated and fluorescence minus-one controls are defined as 100% and 0% staining within each replicate ( n = 3–4 biologically independent replicates). Cathepsins have been labeled with their letter, for example, “A” refers to cathepsin A. CTSE was excluded because of the low pH of the CTSE activation buffer causing cellular toxicity . See for representative flow cytometry histograms, assays performed at pH 5 and 7, and bar graphs with raw median fluorescence intensity (MFI) values. Data are shown as mean ± SD from three to four biologically independent replicates, and each dot represents a single replicate. p Values determined via one-way ANOVA corrected via Dunnett’s multiple comparison test, with a single pooled variance. ∗ p ≤ 0.05, ∗∗ p ≤ 0.01, ∗∗∗ ≤ 0.001, and ∗∗∗∗ p ≤ 0.0001. CTSE, cathepsin E; StcE E447D , inactive point mutant of StcE used as a pan-mucin probe.

Article Snippet: Mucin and MUC1 staining was performed with 15 μg/ml biotin-StcE E447D and 2 μg/ml anti-MUC1 antibody (Novus Bio, clone 967) in PBST for 1 h at room temperature, followed by 3 × 10 min wash with PBS-T. Then, the membrane was incubated with IRDye 800CW Streptavidin and 680RD Goat anti-Mouse IgG (LI-COR Biosciences) according to the manufacturer's recommendations, with 3 × 10 min wash with PBS-T before imaging.

Techniques: Flow Cytometry, Staining, Fluorescence, Labeling, Activation Assay, Comparison, Mutagenesis

Journal: The Journal of Biological Chemistry

Article Title: The protease cathepsin K can debulk the cancer glycocalyx

doi: 10.1016/j.jbc.2026.111206

Figure Lengend Snippet: CTSK degrades cell-surface mucins across multiple cell lines. A, schematic describing the flow cytometry assay for detecting the degradation of cell-surface mucins on H82, OVCAR3, MCF10A, and MCF10 MUC1 cells. B, median fluorescence intensity (MFI) of MUC1 and total mucins via StcE E447D . C, normalized MFI of MUC1 staining for MCF10 ± MUC1 following enzymatic treatments for 1 h a 37 °C at pH 6.75 in Hanks' balanced salt solution (HBSS). D, normalized StcE E447D staining for cell lines following enzymatic treatment for 1 h a 37 °C at pH 6.75. See , A – D for representative histograms. Data are shown as mean ± SD from two to four biologically independent replicates, and each dot represents a single replicate. P Values determined via ( C ) two-way ANOVA or ( D ) one-way ANOVA, both corrected via Tukey’s multiple comparison test, with a single pooled variance for each cell line. ∗ p ≤ 0.05, ∗∗ p ≤ 0.01, ∗∗∗ ≤ 0.001, and ∗∗∗∗ p ≤ 0.0001. CTSK, cathepsin K; MUC, mucin; StcE E447D , inactive point mutant of StcE used as a pan-mucin probe.

Article Snippet: Mucin and MUC1 staining was performed with 15 μg/ml biotin-StcE E447D and 2 μg/ml anti-MUC1 antibody (Novus Bio, clone 967) in PBST for 1 h at room temperature, followed by 3 × 10 min wash with PBS-T. Then, the membrane was incubated with IRDye 800CW Streptavidin and 680RD Goat anti-Mouse IgG (LI-COR Biosciences) according to the manufacturer's recommendations, with 3 × 10 min wash with PBS-T before imaging.

Techniques: Flow Cytometry, Fluorescence, Staining, Comparison, Mutagenesis

Journal: The Journal of Biological Chemistry

Article Title: The protease cathepsin K can debulk the cancer glycocalyx

doi: 10.1016/j.jbc.2026.111206

Figure Lengend Snippet: Cathepsin K (CTSK) tolerates glycans near the cleavage site. A, cleavage motif of CTSK was generated from mass spectrometry analysis of ( left ) glycopeptides, ( center ) nonmodified peptides, and ( right ) modified and nonmodified peptides generated from CTSK digestion of purified and recombinant mucins and nonmucin glycoproteins, followed by trypsin digestion (see the section). The bar graphs on top of the glycopeptide cleavage motif indicate the frequency of O -glycosylation at each threonine and serine residue at that position. B, top, visualization of CTSK cleavage sites in recombinant MUC1 residues 24–47. The sialylated core-1 glycan at specific resides indicates that glycans were seen at those sites. This specific glycan was seen often in the dataset, but its depiction here is only intended to indicate glycosites, not to represent the diversity of all glycans detected at each glycosite in the dataset. Purple diamond , sialic acid; yellow circle , galactose; yellow square , N-acetylgalactosamine; and yellow circle , glycosylation site. Colored bars represent individual detected peptide sequences from CTSK cleavage only, with any shared peptides with chymotrypsin removed. Bottom, annotated spectrum for the indicated MUC1 O -glycopeptide. C, top, visualization of CTSK cleavage sites in recombinant P-selectin glycoprotein ligand-1 (PSGL-1) residues 148 to 197, represented as in ( B ), but this time from the tryptic + CTSK dataset, with all tryptic cleavage sites removed. Bottom, annotated spectrum for the indicated PSGL-1 O -glycopeptide. MUC, mucin.

Article Snippet: Mucin and MUC1 staining was performed with 15 μg/ml biotin-StcE E447D and 2 μg/ml anti-MUC1 antibody (Novus Bio, clone 967) in PBST for 1 h at room temperature, followed by 3 × 10 min wash with PBS-T. Then, the membrane was incubated with IRDye 800CW Streptavidin and 680RD Goat anti-Mouse IgG (LI-COR Biosciences) according to the manufacturer's recommendations, with 3 × 10 min wash with PBS-T before imaging.

Techniques: Generated, Mass Spectrometry, Modification, Purification, Recombinant, Glycoproteomics, Residue

Journal: The Journal of Biological Chemistry

Article Title: The protease cathepsin K can debulk the cancer glycocalyx

doi: 10.1016/j.jbc.2026.111206

Figure Lengend Snippet: Cathepsin K (CTSK) sheds bulky glycan polymers across multiple cell lines. A, schematic describing the flow cytometry assay for detecting shedding of glycan polymers from H82, OVCAR3, MCF10A, and MCF10 MUC1 cells following enzymatic treatments with CTSK, heat-inactivated CTSK (HI CTSK), StcE, heparinase, chondroitinase, and the polySia-specific endosialidase (EndoNA). Cells were stained for ( B ) heparan sulfate using fibroblast growth factor 2 (FGF2), which is a probe for heparan sulfate, ( C ) polysialic acid using anti-polySia antibody (clone 735), ( D ) chondroitin sulfate using anti–chondroitin sulfate antibody (clone CS-56), and ( E ) viability following enzymatic treatment of cells for 1 h at 37 °C at pH 6.75. Staining was ( B ) normalized median fluorescence intensity (MFI) to 100% buffer control and 0% secondary only or ( C and D ) quantified as percent positive staining because of the broad and non-normal distribution of the cell populations. See for normalized staining values and representative histograms. F, change in glycocalyx thickness of YSCCC, MCF10A, and MCF10 MUC1 cells following enzymatic treatment relative to buffer control was measured using scanning angle interference microscopy. Each data point is the average of 20 to 50 individual cell measurements performed on a single day and represents an independent biological replicate. See for individual cell measurements. Data are shown as mean ± SD from two to five biologically independent replicates, and each dot represents a single replicate. p Values determined via one-way or two-way ANOVA corrected via Tukey’s multiple comparison test, with a single pooled variance for each cell line. ∗ p ≤ 0.05, ∗∗ p ≤ 0.01, ∗∗∗ ≤ 0.001, and ∗∗∗∗ p ≤ 0.0001.

Article Snippet: Mucin and MUC1 staining was performed with 15 μg/ml biotin-StcE E447D and 2 μg/ml anti-MUC1 antibody (Novus Bio, clone 967) in PBST for 1 h at room temperature, followed by 3 × 10 min wash with PBS-T. Then, the membrane was incubated with IRDye 800CW Streptavidin and 680RD Goat anti-Mouse IgG (LI-COR Biosciences) according to the manufacturer's recommendations, with 3 × 10 min wash with PBS-T before imaging.

Techniques: Glycoproteomics, Flow Cytometry, Staining, Fluorescence, Control, Microscopy, Comparison

Journal: Clinical and Translational Gastroenterology

Article Title: Mucin 5AC as a Biomarker for Sessile Serrated Lesions: Results From a Systematic Review and Meta-Analysis

doi: 10.14309/ctg.0000000000000831

Figure Lengend Snippet: Expression of MUC5AC in a sessile serrated lesion without dysplasia from the ascending colon. The lesion was stained with MUC5AC ( a and d ), MUC1 ( b and e ), and P40 ( c and f ) and photographed at ×100 ( a , b , and c ) and ×40 ( d , e , and f ). MUC5AC was expressed in serrated glands (*) but not in nearby normal glands (**) ( a and d ). MUC1 was expressed in both serrated glands and normal glands ( b and d ). P40 was not expressed in either serrated or normal glands.

Article Snippet: Histopathologic analysis was performed by 1 author (Z.P.), using monoclonal antibodies against MUC1 (SPM533; Novus Biologicals, Centennial, CO), MUC5AC (917 + 45M1; Novus Biologicals), and P40 (a squamous cell marker) used as a negative control.

Techniques: Expressing, Staining