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mouse monoclonal anti human cd74  (Novus Biologicals)


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    Novus Biologicals mouse monoclonal anti human cd74
    (a) Five melanoma cell lines were analyzed by Western blot analysis for <t>CD74</t> expression. (b) Representative immunohistochemical staining intensities: score 0 (negative), 1 (weak), 2 (medium) and 3 (strong). Scale bar = 100 µm. (c) TMA analysis for CD74 expression during melanoma progression. CD74 staining was scored for the percentage (upper) and intensity (lower) of positive tumor cells. n indicates the number of tissue cores in each group. LN, lymph node. Both percentage and intensity score of each melanoma group were significantly higher than those for nevus group ( P < 0.0001).
    Mouse Monoclonal Anti Human Cd74, supplied by Novus Biologicals, used in various techniques. Bioz Stars score: 93/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/mouse+monoclonal+anti+human+cd74/Human+CD74+Antibody/pmc04640965-131-9-14
    Average 93 stars, based on 6 article reviews
    mouse monoclonal anti human cd74 - by Bioz Stars, 2026-09
    93/100 stars

    Images

    1) Product Images from "Cell Surface CD74-MIF Interactions Drive Melanoma Survival in Response to Interferon-γ"

    Article Title: Cell Surface CD74-MIF Interactions Drive Melanoma Survival in Response to Interferon-γ

    Journal: The Journal of investigative dermatology

    doi: 10.1038/jid.2015.204

    (a) Five melanoma cell lines were analyzed by Western blot analysis for CD74 expression. (b) Representative immunohistochemical staining intensities: score 0 (negative), 1 (weak), 2 (medium) and 3 (strong). Scale bar = 100 µm. (c) TMA analysis for CD74 expression during melanoma progression. CD74 staining was scored for the percentage (upper) and intensity (lower) of positive tumor cells. n indicates the number of tissue cores in each group. LN, lymph node. Both percentage and intensity score of each melanoma group were significantly higher than those for nevus group ( P < 0.0001).
    Figure Legend Snippet: (a) Five melanoma cell lines were analyzed by Western blot analysis for CD74 expression. (b) Representative immunohistochemical staining intensities: score 0 (negative), 1 (weak), 2 (medium) and 3 (strong). Scale bar = 100 µm. (c) TMA analysis for CD74 expression during melanoma progression. CD74 staining was scored for the percentage (upper) and intensity (lower) of positive tumor cells. n indicates the number of tissue cores in each group. LN, lymph node. Both percentage and intensity score of each melanoma group were significantly higher than those for nevus group ( P < 0.0001).

    Techniques Used: Western Blot, Expressing, Immunohistochemical staining, Staining

    (a, b and c) Melanoma cell lines were treated with 0, 100 or 500 IU/ml IFN-γ for 48 hours and then assessed for CD74 expression. (a) Western blot analysis for total CD74 protein. Numbers above each band indicate relative CD74 expression levels. (b) Flow cytometric analysis for cell surface CD74. Histogram: black, isotype control; red, untreated cells stained with FITC-CD74 antibody; green, 100 IU/ml IFN-γ–treated cells with FITC-CD74 antibody; blue, 500 IU/ml IFN-γ–treated cells with FITC-CD74 antibody. (c) Immunofluorescence confocal microscopic analysis of cell surface CD74. Green (Alexa-488), CD74. Blue (DAPI), nucleus. A375 and SB2 cells showed slight expression under normal conditions (arrows). Scale bar = 30 µm. (d) Representative images of weak (left panels, score 1), medium (middle panels, score 2) and strong CD74 staining (right panels, score 3). Upper panel, Scale bar = 1 mm.; lower panel, Scale bar = 250 µm. Corresponding plasma IFN-γ levels are listed. (e) The association between CD74 staining at the tumor site and plasma IFN-γ levels in 55 patients with melanoma. Red bars, means. Error bars, standard deviations. ** P < 0.01, *** P < 0.001.
    Figure Legend Snippet: (a, b and c) Melanoma cell lines were treated with 0, 100 or 500 IU/ml IFN-γ for 48 hours and then assessed for CD74 expression. (a) Western blot analysis for total CD74 protein. Numbers above each band indicate relative CD74 expression levels. (b) Flow cytometric analysis for cell surface CD74. Histogram: black, isotype control; red, untreated cells stained with FITC-CD74 antibody; green, 100 IU/ml IFN-γ–treated cells with FITC-CD74 antibody; blue, 500 IU/ml IFN-γ–treated cells with FITC-CD74 antibody. (c) Immunofluorescence confocal microscopic analysis of cell surface CD74. Green (Alexa-488), CD74. Blue (DAPI), nucleus. A375 and SB2 cells showed slight expression under normal conditions (arrows). Scale bar = 30 µm. (d) Representative images of weak (left panels, score 1), medium (middle panels, score 2) and strong CD74 staining (right panels, score 3). Upper panel, Scale bar = 1 mm.; lower panel, Scale bar = 250 µm. Corresponding plasma IFN-γ levels are listed. (e) The association between CD74 staining at the tumor site and plasma IFN-γ levels in 55 patients with melanoma. Red bars, means. Error bars, standard deviations. ** P < 0.01, *** P < 0.001.

    Techniques Used: Expressing, Western Blot, Control, Staining, Immunofluorescence, Clinical Proteomics

    (a and b) Cells were treated with ISO-1 for 48 hours and then analyzed by Western blot analysis for AKT Ser473 phosphorylation (a) and BCL-2 expression (b). Numbers above each band indicate the relative level of phosphorylated AKT Ser473 (pAKT) to total AKT (AKT) or BCL-2 to actin. (c, d and e) mRNA expression levels of BCL-2 (c), IL-6 (d) and IL-8 (e) in melanoma cells treated with ISO-1 for 24 hours were measured using qRT-PCR. (f) CD74 knockdown A375 cells were analyzed for mRNA expression of the indicated genes by qRT-PCR. shNT, non-target shRNA. * P < 0.05, ** P < 0.01.
    Figure Legend Snippet: (a and b) Cells were treated with ISO-1 for 48 hours and then analyzed by Western blot analysis for AKT Ser473 phosphorylation (a) and BCL-2 expression (b). Numbers above each band indicate the relative level of phosphorylated AKT Ser473 (pAKT) to total AKT (AKT) or BCL-2 to actin. (c, d and e) mRNA expression levels of BCL-2 (c), IL-6 (d) and IL-8 (e) in melanoma cells treated with ISO-1 for 24 hours were measured using qRT-PCR. (f) CD74 knockdown A375 cells were analyzed for mRNA expression of the indicated genes by qRT-PCR. shNT, non-target shRNA. * P < 0.05, ** P < 0.01.

    Techniques Used: Western Blot, Phospho-proteomics, Expressing, Quantitative RT-PCR, Knockdown, shRNA

    Cell surface CD74-negative MeWo cells were subcutaneously injected into the flank of SCID Beige mice. Six days after cell injection, mice were randomly assigned to treatment groups (6 mice per group) and started on daily intraperitoneal treatment with ISO-1 (500 µg/mouse) and/or IFN-γ (1000 IU/mouse). ISO-1 and IFN-γ were administered for 21 days, and tumor growth was monitored. (a) Fold changes of tumor size. (b) Tumor weight on day 21. Error bars, standard deviations. * P < 0.05. (c) Representative mouse from each group (day 21). (d) Immunohistochemical staining for CD74 (upper) and MIF (lower) on day 21. Scale bar = 100 µm. ; AEC chromogen counterstained with hematoxylin.
    Figure Legend Snippet: Cell surface CD74-negative MeWo cells were subcutaneously injected into the flank of SCID Beige mice. Six days after cell injection, mice were randomly assigned to treatment groups (6 mice per group) and started on daily intraperitoneal treatment with ISO-1 (500 µg/mouse) and/or IFN-γ (1000 IU/mouse). ISO-1 and IFN-γ were administered for 21 days, and tumor growth was monitored. (a) Fold changes of tumor size. (b) Tumor weight on day 21. Error bars, standard deviations. * P < 0.05. (c) Representative mouse from each group (day 21). (d) Immunohistochemical staining for CD74 (upper) and MIF (lower) on day 21. Scale bar = 100 µm. ; AEC chromogen counterstained with hematoxylin.

    Techniques Used: Injection, Immunohistochemical staining, Staining

    Related Articles

    Western Blot:

    Article Title: Cell Surface CD74-MIF Interactions Drive Melanoma Survival in Response to Interferon-γ
    Article Snippet: Goat polyclonal anti-human actin (I-19; Santa Cruz, Carlsbad, CA), mouse monoclonal anti-human CD74 (PIN.1; Novus Biologicals, Littleton, CO), mouse monoclonal anti-human BCL-2 (124; GeneTex, San Antonio, TX), goat polyclonal anti-human MIF (R&D Systems, Minneapolis, MN) and rabbit monoclonal anti-human total AKT, phosphorylated AKT Ser473 (Cell Signaling Technology, Danvers, MA) antibodies were used as primary antibodies; horseradish peroxidase-labeled anti-mouse, anti-rabbit and anti-goat antibodies (1:1000; DAKO) were used as secondary antibodies.. Amersham ECL Western blot detection reagent (GE Healthcare, Piscataway, NJ) was used to detect protein expression.Amersham ECL Western blot detection reagent (GE Healthcare, Piscataway, NJ) was used to detect protein expression.

    Expressing:

    Article Title: Cell Surface CD74-MIF Interactions Drive Melanoma Survival in Response to Interferon-γ
    Article Snippet: Goat polyclonal anti-human actin (I-19; Santa Cruz, Carlsbad, CA), mouse monoclonal anti-human CD74 (PIN.1; Novus Biologicals, Littleton, CO), mouse monoclonal anti-human BCL-2 (124; GeneTex, San Antonio, TX), goat polyclonal anti-human MIF (R&D Systems, Minneapolis, MN) and rabbit monoclonal anti-human total AKT, phosphorylated AKT Ser473 (Cell Signaling Technology, Danvers, MA) antibodies were used as primary antibodies; horseradish peroxidase-labeled anti-mouse, anti-rabbit and anti-goat antibodies (1:1000; DAKO) were used as secondary antibodies.. Amersham ECL Western blot detection reagent (GE Healthcare, Piscataway, NJ) was used to detect protein expression.Amersham ECL Western blot detection reagent (GE Healthcare, Piscataway, NJ) was used to detect protein expression.

    Immunohistochemical staining:

    Article Title: Cell Surface CD74-MIF Interactions Drive Melanoma Survival in Response to Interferon-γ
    Article Snippet: Goat polyclonal anti-human actin (I-19; Santa Cruz, Carlsbad, CA), mouse monoclonal anti-human CD74 (PIN.1; Novus Biologicals, Littleton, CO), mouse monoclonal anti-human BCL-2 (124; GeneTex, San Antonio, TX), goat polyclonal anti-human MIF (R&D Systems, Minneapolis, MN) and rabbit monoclonal anti-human total AKT, phosphorylated AKT Ser473 (Cell Signaling Technology, Danvers, MA) antibodies were used as primary antibodies; horseradish peroxidase-labeled anti-mouse, anti-rabbit and anti-goat antibodies (1:1000; DAKO) were used as secondary antibodies.. Amersham ECL Western blot detection reagent (GE Healthcare, Piscataway, NJ) was used to detect protein expression.Amersham ECL Western blot detection reagent (GE Healthcare, Piscataway, NJ) was used to detect protein expression.

    Staining:

    Article Title: Cell Surface CD74-MIF Interactions Drive Melanoma Survival in Response to Interferon-γ
    Article Snippet: Goat polyclonal anti-human actin (I-19; Santa Cruz, Carlsbad, CA), mouse monoclonal anti-human CD74 (PIN.1; Novus Biologicals, Littleton, CO), mouse monoclonal anti-human BCL-2 (124; GeneTex, San Antonio, TX), goat polyclonal anti-human MIF (R&D Systems, Minneapolis, MN) and rabbit monoclonal anti-human total AKT, phosphorylated AKT Ser473 (Cell Signaling Technology, Danvers, MA) antibodies were used as primary antibodies; horseradish peroxidase-labeled anti-mouse, anti-rabbit and anti-goat antibodies (1:1000; DAKO) were used as secondary antibodies.. Amersham ECL Western blot detection reagent (GE Healthcare, Piscataway, NJ) was used to detect protein expression.Amersham ECL Western blot detection reagent (GE Healthcare, Piscataway, NJ) was used to detect protein expression.

    Control:

    Article Title: Cell Surface CD74-MIF Interactions Drive Melanoma Survival in Response to Interferon-γ
    Article Snippet: Goat polyclonal anti-human actin (I-19; Santa Cruz, Carlsbad, CA), mouse monoclonal anti-human CD74 (PIN.1; Novus Biologicals, Littleton, CO), mouse monoclonal anti-human BCL-2 (124; GeneTex, San Antonio, TX), goat polyclonal anti-human MIF (R&D Systems, Minneapolis, MN) and rabbit monoclonal anti-human total AKT, phosphorylated AKT Ser473 (Cell Signaling Technology, Danvers, MA) antibodies were used as primary antibodies; horseradish peroxidase-labeled anti-mouse, anti-rabbit and anti-goat antibodies (1:1000; DAKO) were used as secondary antibodies.. Amersham ECL Western blot detection reagent (GE Healthcare, Piscataway, NJ) was used to detect protein expression.Amersham ECL Western blot detection reagent (GE Healthcare, Piscataway, NJ) was used to detect protein expression.

    Immunofluorescence:

    Article Title: Cell Surface CD74-MIF Interactions Drive Melanoma Survival in Response to Interferon-γ
    Article Snippet: Goat polyclonal anti-human actin (I-19; Santa Cruz, Carlsbad, CA), mouse monoclonal anti-human CD74 (PIN.1; Novus Biologicals, Littleton, CO), mouse monoclonal anti-human BCL-2 (124; GeneTex, San Antonio, TX), goat polyclonal anti-human MIF (R&D Systems, Minneapolis, MN) and rabbit monoclonal anti-human total AKT, phosphorylated AKT Ser473 (Cell Signaling Technology, Danvers, MA) antibodies were used as primary antibodies; horseradish peroxidase-labeled anti-mouse, anti-rabbit and anti-goat antibodies (1:1000; DAKO) were used as secondary antibodies.. Amersham ECL Western blot detection reagent (GE Healthcare, Piscataway, NJ) was used to detect protein expression.Amersham ECL Western blot detection reagent (GE Healthcare, Piscataway, NJ) was used to detect protein expression.

    Clinical Proteomics:

    Article Title: Cell Surface CD74-MIF Interactions Drive Melanoma Survival in Response to Interferon-γ
    Article Snippet: Goat polyclonal anti-human actin (I-19; Santa Cruz, Carlsbad, CA), mouse monoclonal anti-human CD74 (PIN.1; Novus Biologicals, Littleton, CO), mouse monoclonal anti-human BCL-2 (124; GeneTex, San Antonio, TX), goat polyclonal anti-human MIF (R&D Systems, Minneapolis, MN) and rabbit monoclonal anti-human total AKT, phosphorylated AKT Ser473 (Cell Signaling Technology, Danvers, MA) antibodies were used as primary antibodies; horseradish peroxidase-labeled anti-mouse, anti-rabbit and anti-goat antibodies (1:1000; DAKO) were used as secondary antibodies.. Amersham ECL Western blot detection reagent (GE Healthcare, Piscataway, NJ) was used to detect protein expression.Amersham ECL Western blot detection reagent (GE Healthcare, Piscataway, NJ) was used to detect protein expression.

    Phospho-proteomics:

    Article Title: Cell Surface CD74-MIF Interactions Drive Melanoma Survival in Response to Interferon-γ
    Article Snippet: Goat polyclonal anti-human actin (I-19; Santa Cruz, Carlsbad, CA), mouse monoclonal anti-human CD74 (PIN.1; Novus Biologicals, Littleton, CO), mouse monoclonal anti-human BCL-2 (124; GeneTex, San Antonio, TX), goat polyclonal anti-human MIF (R&D Systems, Minneapolis, MN) and rabbit monoclonal anti-human total AKT, phosphorylated AKT Ser473 (Cell Signaling Technology, Danvers, MA) antibodies were used as primary antibodies; horseradish peroxidase-labeled anti-mouse, anti-rabbit and anti-goat antibodies (1:1000; DAKO) were used as secondary antibodies.. Amersham ECL Western blot detection reagent (GE Healthcare, Piscataway, NJ) was used to detect protein expression.Amersham ECL Western blot detection reagent (GE Healthcare, Piscataway, NJ) was used to detect protein expression.

    Quantitative RT-PCR:

    Article Title: Cell Surface CD74-MIF Interactions Drive Melanoma Survival in Response to Interferon-γ
    Article Snippet: Goat polyclonal anti-human actin (I-19; Santa Cruz, Carlsbad, CA), mouse monoclonal anti-human CD74 (PIN.1; Novus Biologicals, Littleton, CO), mouse monoclonal anti-human BCL-2 (124; GeneTex, San Antonio, TX), goat polyclonal anti-human MIF (R&D Systems, Minneapolis, MN) and rabbit monoclonal anti-human total AKT, phosphorylated AKT Ser473 (Cell Signaling Technology, Danvers, MA) antibodies were used as primary antibodies; horseradish peroxidase-labeled anti-mouse, anti-rabbit and anti-goat antibodies (1:1000; DAKO) were used as secondary antibodies.. Amersham ECL Western blot detection reagent (GE Healthcare, Piscataway, NJ) was used to detect protein expression.Amersham ECL Western blot detection reagent (GE Healthcare, Piscataway, NJ) was used to detect protein expression.

    Knockdown:

    Article Title: Cell Surface CD74-MIF Interactions Drive Melanoma Survival in Response to Interferon-γ
    Article Snippet: Goat polyclonal anti-human actin (I-19; Santa Cruz, Carlsbad, CA), mouse monoclonal anti-human CD74 (PIN.1; Novus Biologicals, Littleton, CO), mouse monoclonal anti-human BCL-2 (124; GeneTex, San Antonio, TX), goat polyclonal anti-human MIF (R&D Systems, Minneapolis, MN) and rabbit monoclonal anti-human total AKT, phosphorylated AKT Ser473 (Cell Signaling Technology, Danvers, MA) antibodies were used as primary antibodies; horseradish peroxidase-labeled anti-mouse, anti-rabbit and anti-goat antibodies (1:1000; DAKO) were used as secondary antibodies.. Amersham ECL Western blot detection reagent (GE Healthcare, Piscataway, NJ) was used to detect protein expression.Amersham ECL Western blot detection reagent (GE Healthcare, Piscataway, NJ) was used to detect protein expression.

    shRNA:

    Article Title: Cell Surface CD74-MIF Interactions Drive Melanoma Survival in Response to Interferon-γ
    Article Snippet: Goat polyclonal anti-human actin (I-19; Santa Cruz, Carlsbad, CA), mouse monoclonal anti-human CD74 (PIN.1; Novus Biologicals, Littleton, CO), mouse monoclonal anti-human BCL-2 (124; GeneTex, San Antonio, TX), goat polyclonal anti-human MIF (R&D Systems, Minneapolis, MN) and rabbit monoclonal anti-human total AKT, phosphorylated AKT Ser473 (Cell Signaling Technology, Danvers, MA) antibodies were used as primary antibodies; horseradish peroxidase-labeled anti-mouse, anti-rabbit and anti-goat antibodies (1:1000; DAKO) were used as secondary antibodies.. Amersham ECL Western blot detection reagent (GE Healthcare, Piscataway, NJ) was used to detect protein expression.Amersham ECL Western blot detection reagent (GE Healthcare, Piscataway, NJ) was used to detect protein expression.

    Injection:

    Article Title: Cell Surface CD74-MIF Interactions Drive Melanoma Survival in Response to Interferon-γ
    Article Snippet: Goat polyclonal anti-human actin (I-19; Santa Cruz, Carlsbad, CA), mouse monoclonal anti-human CD74 (PIN.1; Novus Biologicals, Littleton, CO), mouse monoclonal anti-human BCL-2 (124; GeneTex, San Antonio, TX), goat polyclonal anti-human MIF (R&D Systems, Minneapolis, MN) and rabbit monoclonal anti-human total AKT, phosphorylated AKT Ser473 (Cell Signaling Technology, Danvers, MA) antibodies were used as primary antibodies; horseradish peroxidase-labeled anti-mouse, anti-rabbit and anti-goat antibodies (1:1000; DAKO) were used as secondary antibodies.. Amersham ECL Western blot detection reagent (GE Healthcare, Piscataway, NJ) was used to detect protein expression.Amersham ECL Western blot detection reagent (GE Healthcare, Piscataway, NJ) was used to detect protein expression.



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    Thermo Fisher mouse monoclonal anti-human cd74
    Human Colonic Mesenchymal Heterogeneity in Health (A) Flow cytometry analysis of the indicated surface markers on colonic single-cell suspensions following removal of epithelial and hematopoietic cells by MACS. Column flow-through is shown in red, and column-retained fraction is in blue. (B) t-SNE plot of the healthy human colonic mesenchyme dataset. Single cells colored by cluster annotation. (C) Violin plots for pan-fibroblast marker genes vimentin ( VIM ) and collagen types 1 and 3 ( COL1A2 , COL3A1 ) across clusters. (D) Violin plots for high-ranked transcriptional regulators and marker genes sharing GO annotation for significantly enriched terms for (i) S1 subset, <t>(ii)</t> S2 subset, (iii) S3 subset, (iv) S4 subset, and (v) myofibroblasts. Crossbars indicate median expression. (E) Single-molecule ISH staining of healthy human colonic tissue showing distribution of S1 markers ( ADAMDEC1 , DCN , SLIT2 , and CXCL12 ) (left) and S2 markers ( F3 (CD142) , WNT5A , HSD17B2 , WNT5B , POSTN , BMP2 , FRZB , BMP5 ) (right). (F) Identification of SOX6 − ZEB2 + /ZEB1 − ZEB2 + S1 and SOX6 + ZEB2 − /ZEB1 + ZEB2 − S2 subsets in healthy human colon. (G) Single (left) and co-staining with CD45 (right) and F3/CD142 (S2), ZEB2 (S1), and SMAD7 (S3) by IHC in colonic sections. The lower far-right panel is a quadruple stain of all 4 markers. (H) Differential expression analysis between S2a and S2b reveals 302 differentially expressed genes. (I) t-SNE plots showing examples of genes differentially expressed between S2a and S2b. (J) GO enrichment terms for S2a and S2b. See also – and – .
    Mouse Monoclonal Anti Human Cd74, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    mouse monoclonal anti human cd74  (Novus Biologicals)
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    Novus Biologicals mouse monoclonal anti human cd74
    (a) Five melanoma cell lines were analyzed by Western blot analysis for <t>CD74</t> expression. (b) Representative immunohistochemical staining intensities: score 0 (negative), 1 (weak), 2 (medium) and 3 (strong). Scale bar = 100 µm. (c) TMA analysis for CD74 expression during melanoma progression. CD74 staining was scored for the percentage (upper) and intensity (lower) of positive tumor cells. n indicates the number of tissue cores in each group. LN, lymph node. Both percentage and intensity score of each melanoma group were significantly higher than those for nevus group ( P < 0.0001).
    Mouse Monoclonal Anti Human Cd74, supplied by Novus Biologicals, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    mouse monoclonal anti-human cd74  (Santa Cruz Biotechnology)
    90
    Santa Cruz Biotechnology mouse monoclonal anti-human cd74
    (a) Five melanoma cell lines were analyzed by Western blot analysis for <t>CD74</t> expression. (b) Representative immunohistochemical staining intensities: score 0 (negative), 1 (weak), 2 (medium) and 3 (strong). Scale bar = 100 µm. (c) TMA analysis for CD74 expression during melanoma progression. CD74 staining was scored for the percentage (upper) and intensity (lower) of positive tumor cells. n indicates the number of tissue cores in each group. LN, lymph node. Both percentage and intensity score of each melanoma group were significantly higher than those for nevus group ( P < 0.0001).
    Mouse Monoclonal Anti Human Cd74, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    mouse monoclonal antibodies  (R&D Systems)
    80
    R&D Systems mouse monoclonal antibodies
    (a) Five melanoma cell lines were analyzed by Western blot analysis for <t>CD74</t> expression. (b) Representative immunohistochemical staining intensities: score 0 (negative), 1 (weak), 2 (medium) and 3 (strong). Scale bar = 100 µm. (c) TMA analysis for CD74 expression during melanoma progression. CD74 staining was scored for the percentage (upper) and intensity (lower) of positive tumor cells. n indicates the number of tissue cores in each group. LN, lymph node. Both percentage and intensity score of each melanoma group were significantly higher than those for nevus group ( P < 0.0001).
    Mouse Monoclonal Antibodies, supplied by R&D Systems, used in various techniques. Bioz Stars score: 80/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    T cell and microbial co-localization define distinct cellular neighborhoods in human pancreatic cancer (A) Study design for (B)–(D) (n = 185 for cold nests and n = 77 for hot nests pooled from 16 specimens). (B) Images of CD8 (yellow), CD68 (brown), tumor epithelium (teal, CK19), FOXP3 (purple), and nuclei (blue, hematoxylin). Scale bars, 1 mm (top) and 50 μm (bottom). (C) Quantification of CD8 + (left), FOXP3 + (middle), and CD68 + (right) cells. (D) Ratio of CD8 + cells to FOXP3 + cells. (E) Study design for (F) and (H)–(J). (F) Principal-component analysis (n = 5 for hot and cold tumor nests from 1 specimen). (G) 16S rRNA levels (n = 14 for cold and hot tumor nests from 3 patient specimens). (H and I) Heatmap (H) and enrichment score (I) for the relative expression of genes associated with response to bacterium (GO: 0009617) in hot and cold stroma. (J) Volcano plot of DEGs. (K) Multiplex immunohistochemistry images of CD8 (yellow), PIGR (purple), and nuclei (blue, hematoxylin). (L) Quantification of PIGR (n = 49 for cold nests and n = 46 for hot nests pooled from 7 specimens). (M) Multiplex immunohistochemistry images of CD8 (teal), CD20 (yellow), CD74 (purple), and nuclei (blue, hematoxylin). (N) Quantification of CD74 (n = 71 for cold nests and n = 45 for hot nests pooled from 10 specimens). (O) Summary of cell markers and proteins. For (K) and (M), scale bars, 2 mm (left) and 50 μm (right), and dashed lines indicate tumor epithelium. Statistical significance was calculated using a two-tailed Mann-Whitney test. Data are represented as violin plots (center line, median; top and bottom lines, upper and lower quartiles) and scatterplots (mean + SD or mean). PDAC, pancreatic ductal adenocarcinoma.

    Journal: Cell Reports Medicine

    Article Title: Multimodal immune phenotyping reveals microbial-T cell interactions that shape pancreatic cancer

    doi: 10.1016/j.xcrm.2024.101397

    Figure Lengend Snippet: T cell and microbial co-localization define distinct cellular neighborhoods in human pancreatic cancer (A) Study design for (B)–(D) (n = 185 for cold nests and n = 77 for hot nests pooled from 16 specimens). (B) Images of CD8 (yellow), CD68 (brown), tumor epithelium (teal, CK19), FOXP3 (purple), and nuclei (blue, hematoxylin). Scale bars, 1 mm (top) and 50 μm (bottom). (C) Quantification of CD8 + (left), FOXP3 + (middle), and CD68 + (right) cells. (D) Ratio of CD8 + cells to FOXP3 + cells. (E) Study design for (F) and (H)–(J). (F) Principal-component analysis (n = 5 for hot and cold tumor nests from 1 specimen). (G) 16S rRNA levels (n = 14 for cold and hot tumor nests from 3 patient specimens). (H and I) Heatmap (H) and enrichment score (I) for the relative expression of genes associated with response to bacterium (GO: 0009617) in hot and cold stroma. (J) Volcano plot of DEGs. (K) Multiplex immunohistochemistry images of CD8 (yellow), PIGR (purple), and nuclei (blue, hematoxylin). (L) Quantification of PIGR (n = 49 for cold nests and n = 46 for hot nests pooled from 7 specimens). (M) Multiplex immunohistochemistry images of CD8 (teal), CD20 (yellow), CD74 (purple), and nuclei (blue, hematoxylin). (N) Quantification of CD74 (n = 71 for cold nests and n = 45 for hot nests pooled from 10 specimens). (O) Summary of cell markers and proteins. For (K) and (M), scale bars, 2 mm (left) and 50 μm (right), and dashed lines indicate tumor epithelium. Statistical significance was calculated using a two-tailed Mann-Whitney test. Data are represented as violin plots (center line, median; top and bottom lines, upper and lower quartiles) and scatterplots (mean + SD or mean). PDAC, pancreatic ductal adenocarcinoma.

    Article Snippet: Mouse monoclonal anti-human CD74 (LN2) , Abcam , ab9514; RRID:AB_2075504.

    Techniques: Expressing, Multiplex Assay, Immunohistochemistry, Two Tailed Test, MANN-WHITNEY

    T cell infiltration into tumors occurs independent of the gut and tumor microbiome (A) Study design for (B)–(K). (B) 16S rRNA levels in tumor and stool from mice orthotopically injected with cold (69) tumor cells (n = 10) or hot (2838c3) tumor cells and treated with (n = 15) and without (n = 20) antibiotics. (C and D) Quantification of CD3 + (C) and CD8 + and FOXP3 + (D) T cells from hot tumors of mice treated with (n = 19) or without (n = 13) antibiotics. (E) 16S rRNA levels in tumor from mice orthotopically injected with cold (69) tumor cells and treated with (n = 5) and without (n = 5) antibiotics. (F) Quantification of CD8 + and FOXP3 + T cells from cold tumors of mice treated with (n = 5) and without (n = 5) antibiotics. (G) Number of DEGs. (H) Bar graph displaying overrepresentation analysis of DEGs in indicated gene sets. (I and J) Tumor weights at day 20. (K) Quantification of intra-tumoral CD19 + cells. (L and M) Mean fluorescence intensity (MFI) of MHC class II (L) and CD206 (M) on CD11b + F4/80 + intra-tumoral macrophages. Data were pooled from two to three experiments (B–D and J–M) or are representative of two independent experiments (E–I). Statistical significance was calculated using one-way ANOVA with Dunnett’s test (B, L, and M) and a two-tailed Mann-Whitney test (C–F and I–K). Data are represented as scatterplots (mean ± SD). NS, not significant.

    Journal: Cell Reports Medicine

    Article Title: Multimodal immune phenotyping reveals microbial-T cell interactions that shape pancreatic cancer

    doi: 10.1016/j.xcrm.2024.101397

    Figure Lengend Snippet: T cell infiltration into tumors occurs independent of the gut and tumor microbiome (A) Study design for (B)–(K). (B) 16S rRNA levels in tumor and stool from mice orthotopically injected with cold (69) tumor cells (n = 10) or hot (2838c3) tumor cells and treated with (n = 15) and without (n = 20) antibiotics. (C and D) Quantification of CD3 + (C) and CD8 + and FOXP3 + (D) T cells from hot tumors of mice treated with (n = 19) or without (n = 13) antibiotics. (E) 16S rRNA levels in tumor from mice orthotopically injected with cold (69) tumor cells and treated with (n = 5) and without (n = 5) antibiotics. (F) Quantification of CD8 + and FOXP3 + T cells from cold tumors of mice treated with (n = 5) and without (n = 5) antibiotics. (G) Number of DEGs. (H) Bar graph displaying overrepresentation analysis of DEGs in indicated gene sets. (I and J) Tumor weights at day 20. (K) Quantification of intra-tumoral CD19 + cells. (L and M) Mean fluorescence intensity (MFI) of MHC class II (L) and CD206 (M) on CD11b + F4/80 + intra-tumoral macrophages. Data were pooled from two to three experiments (B–D and J–M) or are representative of two independent experiments (E–I). Statistical significance was calculated using one-way ANOVA with Dunnett’s test (B, L, and M) and a two-tailed Mann-Whitney test (C–F and I–K). Data are represented as scatterplots (mean ± SD). NS, not significant.

    Article Snippet: Mouse monoclonal anti-human CD74 (LN2) , Abcam , ab9514; RRID:AB_2075504.

    Techniques: Injection, Fluorescence, Two Tailed Test, MANN-WHITNEY

    Journal: Cell Reports Medicine

    Article Title: Multimodal immune phenotyping reveals microbial-T cell interactions that shape pancreatic cancer

    doi: 10.1016/j.xcrm.2024.101397

    Figure Lengend Snippet:

    Article Snippet: Mouse monoclonal anti-human CD74 (LN2) , Abcam , ab9514; RRID:AB_2075504.

    Techniques: Recombinant, Sequencing, Software

    LN-2 and UMAb231 antibody specificity. ( A ) Western blot analysis of CD74 expression was performed by loading whole cell extracts of MDA-MB-231 control cells (control), MDA-MD-231 cells with CD74 siRNA knockdown (+CCD74 siRNA), and IFNγ stimulated MDA-MD-231 cells (+IFNγ). CD74 protein was detected at 33/35 kDa using either anti-CD74 LN-2 (left-hand panel) or UMAb231 monoclonal antibody (right-hand panel), and at 41/43 kDa with UMAb231 in overexpressing cells. *, **, and *** mark the positions of three independent bands of unknown identity. No signal was detected in siRNA-mediated CD74 knockdown cells (+CD74 siRNA). IHC analysis of FFPE sections of MDA-MB-231 breast cancer control cells compared to ( B ) CD74 siRNA knockdown, and ( C ) IFNγ stimulation showed similar staining patterns with LN-2 and UMAb231 antibodies. Insets in subpanels 1b and 1c show higher magnification images illustrating punctuated stainings of CD74. ( D ) Quantification of stainings showed that IHC immunoreactivity correlated with expression of cellular CD74 protein. Scale bars = 20 µm. The uncropped blot is shown in .

    Journal: Cancers

    Article Title: HLA Class II Histocompatibility Antigen γ Chain (CD74) Expression Is Associated with Immune Cell Infiltration and Favorable Outcome in Breast Cancer

    doi: 10.3390/cancers13246179

    Figure Lengend Snippet: LN-2 and UMAb231 antibody specificity. ( A ) Western blot analysis of CD74 expression was performed by loading whole cell extracts of MDA-MB-231 control cells (control), MDA-MD-231 cells with CD74 siRNA knockdown (+CCD74 siRNA), and IFNγ stimulated MDA-MD-231 cells (+IFNγ). CD74 protein was detected at 33/35 kDa using either anti-CD74 LN-2 (left-hand panel) or UMAb231 monoclonal antibody (right-hand panel), and at 41/43 kDa with UMAb231 in overexpressing cells. *, **, and *** mark the positions of three independent bands of unknown identity. No signal was detected in siRNA-mediated CD74 knockdown cells (+CD74 siRNA). IHC analysis of FFPE sections of MDA-MB-231 breast cancer control cells compared to ( B ) CD74 siRNA knockdown, and ( C ) IFNγ stimulation showed similar staining patterns with LN-2 and UMAb231 antibodies. Insets in subpanels 1b and 1c show higher magnification images illustrating punctuated stainings of CD74. ( D ) Quantification of stainings showed that IHC immunoreactivity correlated with expression of cellular CD74 protein. Scale bars = 20 µm. The uncropped blot is shown in .

    Article Snippet: Recently, a new anti-CD74 mouse monoclonal antibody, UMAb231, was developed by Origene Technologies (Rockville, MD, USA).

    Techniques: Western Blot, Expressing, Staining

    IHC staining patterns of UMAb231. Immunoreactivity for CD74 was seen primarily in stromal components, ( A ) in peritumoral fibroblasts, ( B ) endothelial cells, and ( C ) lymphocytes and macrophages, respectively (black arrows). Staining of serial sections with vimentin, CD4, CD8, CD68 or CD31 was used to confirm cell identity. In some samples, CD74 was also expressed in ( D ) tumor cells (black arrow indicates tumor cells), with essentially cytoplasmic and ( E ) membrane staining (black arrow indicates membrane staining). ( F ) Comparative semi-quantitative analysis of the UMAb231 and LN-2 antibodies, showed similar, but not always identical, patterns between the two antibodies. A case is highlighted with no CD74 immunoreactivity in tumor cells with UMAb231 (sub-panel b), but weak to moderate reactivity with LN-2 (sub-panel a, black arrow). In both cases lymphocytes and macrophages stained strongly with either antibody (sub-panels a and b, red arrows, respectively). Scale bars are 100 µm in ( A , D , F ), 25 µm in B, 250 µm in ( C ), and 50 µm in ( E ), respectively.

    Journal: Cancers

    Article Title: HLA Class II Histocompatibility Antigen γ Chain (CD74) Expression Is Associated with Immune Cell Infiltration and Favorable Outcome in Breast Cancer

    doi: 10.3390/cancers13246179

    Figure Lengend Snippet: IHC staining patterns of UMAb231. Immunoreactivity for CD74 was seen primarily in stromal components, ( A ) in peritumoral fibroblasts, ( B ) endothelial cells, and ( C ) lymphocytes and macrophages, respectively (black arrows). Staining of serial sections with vimentin, CD4, CD8, CD68 or CD31 was used to confirm cell identity. In some samples, CD74 was also expressed in ( D ) tumor cells (black arrow indicates tumor cells), with essentially cytoplasmic and ( E ) membrane staining (black arrow indicates membrane staining). ( F ) Comparative semi-quantitative analysis of the UMAb231 and LN-2 antibodies, showed similar, but not always identical, patterns between the two antibodies. A case is highlighted with no CD74 immunoreactivity in tumor cells with UMAb231 (sub-panel b), but weak to moderate reactivity with LN-2 (sub-panel a, black arrow). In both cases lymphocytes and macrophages stained strongly with either antibody (sub-panels a and b, red arrows, respectively). Scale bars are 100 µm in ( A , D , F ), 25 µm in B, 250 µm in ( C ), and 50 µm in ( E ), respectively.

    Article Snippet: Recently, a new anti-CD74 mouse monoclonal antibody, UMAb231, was developed by Origene Technologies (Rockville, MD, USA).

    Techniques: Immunohistochemistry, Staining

    IHC analysis of YTMA49. TMA cores were scored in a categorical manner according to CD74 immunoreactivity levels in tumor cells only, as follows: 0 (absent), 1+ (weak), 2+ (moderate) or 3+ (strong) (( A , C , E , G ), respectively). An H-score value was also derived by computer-assisted whole tissue core analysis ( B , D , F , H ). Negative, weak, moderate and strong immunostaining thresholds are yellow, orange, and red, respectively. Intensity scores were correlated to ( I ) TN status and ( J ) node status. Scale bars = 250 µm.

    Journal: Cancers

    Article Title: HLA Class II Histocompatibility Antigen γ Chain (CD74) Expression Is Associated with Immune Cell Infiltration and Favorable Outcome in Breast Cancer

    doi: 10.3390/cancers13246179

    Figure Lengend Snippet: IHC analysis of YTMA49. TMA cores were scored in a categorical manner according to CD74 immunoreactivity levels in tumor cells only, as follows: 0 (absent), 1+ (weak), 2+ (moderate) or 3+ (strong) (( A , C , E , G ), respectively). An H-score value was also derived by computer-assisted whole tissue core analysis ( B , D , F , H ). Negative, weak, moderate and strong immunostaining thresholds are yellow, orange, and red, respectively. Intensity scores were correlated to ( I ) TN status and ( J ) node status. Scale bars = 250 µm.

    Article Snippet: Recently, a new anti-CD74 mouse monoclonal antibody, UMAb231, was developed by Origene Technologies (Rockville, MD, USA).

    Techniques: Derivative Assay, Immunostaining

    Low/high score association with pathology scores, nodal status and TN status (categorical variables).

    Journal: Cancers

    Article Title: HLA Class II Histocompatibility Antigen γ Chain (CD74) Expression Is Associated with Immune Cell Infiltration and Favorable Outcome in Breast Cancer

    doi: 10.3390/cancers13246179

    Figure Lengend Snippet: Low/high score association with pathology scores, nodal status and TN status (categorical variables).

    Article Snippet: Recently, a new anti-CD74 mouse monoclonal antibody, UMAb231, was developed by Origene Technologies (Rockville, MD, USA).

    Techniques:

    Survival analysis of breast cancer samples as a function of CD74 expression. ( A ) The prognostic value of CD74 expression was examined in the YTMA49 cohort for all cases, TN cases, and non-TN cases. ( B ) Kaplan–Meier plot showing OS among breast cancer patients according to CD74 expression in a multivariate model correcting for immune cell infiltration in a multivariable Cox proportional hazard model. Correlation between CD74 gene expression and infiltration of immune cells was examined in the breast cancer samples available from TCGA database with the TIMER analysis tool ( http://timer.cistrome.org ) (accessed on 8 March 2019) ( C ) Correlation between CD74 and PD-L1 in 1017 breast carcinomas from TCGA after correcting for tumor-infiltrating immune cells.

    Journal: Cancers

    Article Title: HLA Class II Histocompatibility Antigen γ Chain (CD74) Expression Is Associated with Immune Cell Infiltration and Favorable Outcome in Breast Cancer

    doi: 10.3390/cancers13246179

    Figure Lengend Snippet: Survival analysis of breast cancer samples as a function of CD74 expression. ( A ) The prognostic value of CD74 expression was examined in the YTMA49 cohort for all cases, TN cases, and non-TN cases. ( B ) Kaplan–Meier plot showing OS among breast cancer patients according to CD74 expression in a multivariate model correcting for immune cell infiltration in a multivariable Cox proportional hazard model. Correlation between CD74 gene expression and infiltration of immune cells was examined in the breast cancer samples available from TCGA database with the TIMER analysis tool ( http://timer.cistrome.org ) (accessed on 8 March 2019) ( C ) Correlation between CD74 and PD-L1 in 1017 breast carcinomas from TCGA after correcting for tumor-infiltrating immune cells.

    Article Snippet: Recently, a new anti-CD74 mouse monoclonal antibody, UMAb231, was developed by Origene Technologies (Rockville, MD, USA).

    Techniques: Expressing

    Human Colonic Mesenchymal Heterogeneity in Health (A) Flow cytometry analysis of the indicated surface markers on colonic single-cell suspensions following removal of epithelial and hematopoietic cells by MACS. Column flow-through is shown in red, and column-retained fraction is in blue. (B) t-SNE plot of the healthy human colonic mesenchyme dataset. Single cells colored by cluster annotation. (C) Violin plots for pan-fibroblast marker genes vimentin ( VIM ) and collagen types 1 and 3 ( COL1A2 , COL3A1 ) across clusters. (D) Violin plots for high-ranked transcriptional regulators and marker genes sharing GO annotation for significantly enriched terms for (i) S1 subset, (ii) S2 subset, (iii) S3 subset, (iv) S4 subset, and (v) myofibroblasts. Crossbars indicate median expression. (E) Single-molecule ISH staining of healthy human colonic tissue showing distribution of S1 markers ( ADAMDEC1 , DCN , SLIT2 , and CXCL12 ) (left) and S2 markers ( F3 (CD142) , WNT5A , HSD17B2 , WNT5B , POSTN , BMP2 , FRZB , BMP5 ) (right). (F) Identification of SOX6 − ZEB2 + /ZEB1 − ZEB2 + S1 and SOX6 + ZEB2 − /ZEB1 + ZEB2 − S2 subsets in healthy human colon. (G) Single (left) and co-staining with CD45 (right) and F3/CD142 (S2), ZEB2 (S1), and SMAD7 (S3) by IHC in colonic sections. The lower far-right panel is a quadruple stain of all 4 markers. (H) Differential expression analysis between S2a and S2b reveals 302 differentially expressed genes. (I) t-SNE plots showing examples of genes differentially expressed between S2a and S2b. (J) GO enrichment terms for S2a and S2b. See also – and – .

    Journal: Cell

    Article Title: Structural Remodeling of the Human Colonic Mesenchyme in Inflammatory Bowel Disease

    doi: 10.1016/j.cell.2018.08.067

    Figure Lengend Snippet: Human Colonic Mesenchymal Heterogeneity in Health (A) Flow cytometry analysis of the indicated surface markers on colonic single-cell suspensions following removal of epithelial and hematopoietic cells by MACS. Column flow-through is shown in red, and column-retained fraction is in blue. (B) t-SNE plot of the healthy human colonic mesenchyme dataset. Single cells colored by cluster annotation. (C) Violin plots for pan-fibroblast marker genes vimentin ( VIM ) and collagen types 1 and 3 ( COL1A2 , COL3A1 ) across clusters. (D) Violin plots for high-ranked transcriptional regulators and marker genes sharing GO annotation for significantly enriched terms for (i) S1 subset, (ii) S2 subset, (iii) S3 subset, (iv) S4 subset, and (v) myofibroblasts. Crossbars indicate median expression. (E) Single-molecule ISH staining of healthy human colonic tissue showing distribution of S1 markers ( ADAMDEC1 , DCN , SLIT2 , and CXCL12 ) (left) and S2 markers ( F3 (CD142) , WNT5A , HSD17B2 , WNT5B , POSTN , BMP2 , FRZB , BMP5 ) (right). (F) Identification of SOX6 − ZEB2 + /ZEB1 − ZEB2 + S1 and SOX6 + ZEB2 − /ZEB1 + ZEB2 − S2 subsets in healthy human colon. (G) Single (left) and co-staining with CD45 (right) and F3/CD142 (S2), ZEB2 (S1), and SMAD7 (S3) by IHC in colonic sections. The lower far-right panel is a quadruple stain of all 4 markers. (H) Differential expression analysis between S2a and S2b reveals 302 differentially expressed genes. (I) t-SNE plots showing examples of genes differentially expressed between S2a and S2b. (J) GO enrichment terms for S2a and S2b. See also – and – .

    Article Snippet: Mouse monoclonal anti-human CD74 , eBioscience , Cat#11-0748-41; RRID: AB_2043845.

    Techniques: Flow Cytometry, Marker, Expressing, Staining, Quantitative Proteomics

    Colonic Mesenchymal Plasticity in IBD (A) t-SNE plot of UC colonic mesenchyme dataset. Single cells colored by cluster annotation. Descriptive cluster labels are shown. (B) Human healthy and UC cluster marker gene overlap correlation heatmap. (C) Selected enriched (FDR < 0.01) GO terms of UC S4 mesenchymal population marker genes. (D) (i) Flow cytometry analysis of CD74 and PDPN expression on colonic stromal cells from Ctrl (right) or UC (left) donors. (ii) Comparison of intracellular CCL19 and IL-33 levels in CD74 high PDPN high CD24 high cells (red) versus the corresponding CD74 low PDPN low subset (blue) in inflamed UC colonic tissue. (E) Flow cytometry analysis of FDCSP high and CD24 high colonic stromal cells from Ctrl (blue) or UC (red). (F) Single-molecule ISH staining of FDCSP in Ctrl or UC colonic tissue sections. (G) Flow cytometric analysis of SOX6 expression in Ctrl (blue) or UC (red) colonic stromal cells. See also and and and .

    Journal: Cell

    Article Title: Structural Remodeling of the Human Colonic Mesenchyme in Inflammatory Bowel Disease

    doi: 10.1016/j.cell.2018.08.067

    Figure Lengend Snippet: Colonic Mesenchymal Plasticity in IBD (A) t-SNE plot of UC colonic mesenchyme dataset. Single cells colored by cluster annotation. Descriptive cluster labels are shown. (B) Human healthy and UC cluster marker gene overlap correlation heatmap. (C) Selected enriched (FDR < 0.01) GO terms of UC S4 mesenchymal population marker genes. (D) (i) Flow cytometry analysis of CD74 and PDPN expression on colonic stromal cells from Ctrl (right) or UC (left) donors. (ii) Comparison of intracellular CCL19 and IL-33 levels in CD74 high PDPN high CD24 high cells (red) versus the corresponding CD74 low PDPN low subset (blue) in inflamed UC colonic tissue. (E) Flow cytometry analysis of FDCSP high and CD24 high colonic stromal cells from Ctrl (blue) or UC (red). (F) Single-molecule ISH staining of FDCSP in Ctrl or UC colonic tissue sections. (G) Flow cytometric analysis of SOX6 expression in Ctrl (blue) or UC (red) colonic stromal cells. See also and and and .

    Article Snippet: Mouse monoclonal anti-human CD74 , eBioscience , Cat#11-0748-41; RRID: AB_2043845.

    Techniques: Marker, Flow Cytometry, Expressing, Comparison, Staining

    Functional Attributes of Crypt Niche and IBD-Associated Mesenchymal Cells (A) Epithelial characterization after in vitro co-culture with and without S2. S2 was isolated by fluorescence-activated cell sorting (FACS) for F3 (CD142). Crypts with (ii) and without (iii) F3 + stromal cells grown in culture containing Rspo1 and assessed for up to 10 days of culture. Representative images from day 4 and day 10 are shown. (i) Normal growth of human colon organoids without any stromal cells. Bar graph shows quantification of organoid complexity during the course of co-culture. (B) (i) Violin plots from the scRNA-seq data showing IL-6 and TNFSF14 (LIGHT) upregulated by S4. (ii) Human colon organoids were treated with 100 ng/mL of either IL-6 or LIGHT. Confocal immunofluorescence images show EdU-labeled nuclei (red) and total nuclei stained with DAPI (blue). Epithelial proliferative capacity was assessed by quantification of the total numbers of EdU positive nuclei and DAPI-stained nuclei to calculate the fraction of proliferating cells in a section of interest. For each experiment, 15 random fields were quantified for each treatment. n = 3 independent experiments. ∗∗ p < 0.0001, ∗ p < 0.001 Mann-Whitney U test. (C) Real-time qPCR measured stem cell markers ( LGR5 , OLFM4 , AXIN2 , NOTCH1 , and ALDH1A1 ) and CDX2 gene expression after treatment of human colon organoids with IL-6 or LIGHT for 4 days in the presence of Wnt containing medium. (D) Real-time qPCR measured stem cell marker ( LGR5 , OLFM4 , AXIN2 , ALDH1A1 , MSI1 , and SOX9 ) and differentiation marker ( KRT20 , MUC2 , and CDX2 ) gene expression after treatment of human colon organoids with IL-6 or LIGHT for 4 days in the presence of Wnt containing medium, with subsequent Wnt withdrawal and treatment with IL-6 and LIGHT for another 4 days. (E) OLFM4 gene expression from scRNA-seq of over 11,175 single cells isolated from healthy, non-inflamed and inflamed colonic biopsies (i), and gene expression from bulk RNA of inflamed and non-inflamed mucosa of IBD patients compared to healthy control samples. (F) (i) Violin plots of relative gene expression of Lox and Loxl1 in DSS-induced colitis. (ii) Cumulative diarrhea score, blood score, and large bowel weight to length ratio of vehicle-only Ctrls versus BAPN-treated animals. (G) Lipid peroxidation measured by malondialdehyde (MDA) plasma levels of vehicle-only and BAPN-treated animals. Error bars represent the SEM.

    Journal: Cell

    Article Title: Structural Remodeling of the Human Colonic Mesenchyme in Inflammatory Bowel Disease

    doi: 10.1016/j.cell.2018.08.067

    Figure Lengend Snippet: Functional Attributes of Crypt Niche and IBD-Associated Mesenchymal Cells (A) Epithelial characterization after in vitro co-culture with and without S2. S2 was isolated by fluorescence-activated cell sorting (FACS) for F3 (CD142). Crypts with (ii) and without (iii) F3 + stromal cells grown in culture containing Rspo1 and assessed for up to 10 days of culture. Representative images from day 4 and day 10 are shown. (i) Normal growth of human colon organoids without any stromal cells. Bar graph shows quantification of organoid complexity during the course of co-culture. (B) (i) Violin plots from the scRNA-seq data showing IL-6 and TNFSF14 (LIGHT) upregulated by S4. (ii) Human colon organoids were treated with 100 ng/mL of either IL-6 or LIGHT. Confocal immunofluorescence images show EdU-labeled nuclei (red) and total nuclei stained with DAPI (blue). Epithelial proliferative capacity was assessed by quantification of the total numbers of EdU positive nuclei and DAPI-stained nuclei to calculate the fraction of proliferating cells in a section of interest. For each experiment, 15 random fields were quantified for each treatment. n = 3 independent experiments. ∗∗ p < 0.0001, ∗ p < 0.001 Mann-Whitney U test. (C) Real-time qPCR measured stem cell markers ( LGR5 , OLFM4 , AXIN2 , NOTCH1 , and ALDH1A1 ) and CDX2 gene expression after treatment of human colon organoids with IL-6 or LIGHT for 4 days in the presence of Wnt containing medium. (D) Real-time qPCR measured stem cell marker ( LGR5 , OLFM4 , AXIN2 , ALDH1A1 , MSI1 , and SOX9 ) and differentiation marker ( KRT20 , MUC2 , and CDX2 ) gene expression after treatment of human colon organoids with IL-6 or LIGHT for 4 days in the presence of Wnt containing medium, with subsequent Wnt withdrawal and treatment with IL-6 and LIGHT for another 4 days. (E) OLFM4 gene expression from scRNA-seq of over 11,175 single cells isolated from healthy, non-inflamed and inflamed colonic biopsies (i), and gene expression from bulk RNA of inflamed and non-inflamed mucosa of IBD patients compared to healthy control samples. (F) (i) Violin plots of relative gene expression of Lox and Loxl1 in DSS-induced colitis. (ii) Cumulative diarrhea score, blood score, and large bowel weight to length ratio of vehicle-only Ctrls versus BAPN-treated animals. (G) Lipid peroxidation measured by malondialdehyde (MDA) plasma levels of vehicle-only and BAPN-treated animals. Error bars represent the SEM.

    Article Snippet: Mouse monoclonal anti-human CD74 , eBioscience , Cat#11-0748-41; RRID: AB_2043845.

    Techniques: Functional Assay, In Vitro, Co-Culture Assay, Isolation, Fluorescence, FACS, Immunofluorescence, Labeling, Staining, MANN-WHITNEY, Gene Expression, Marker, Control, Clinical Proteomics

    Journal: Cell

    Article Title: Structural Remodeling of the Human Colonic Mesenchyme in Inflammatory Bowel Disease

    doi: 10.1016/j.cell.2018.08.067

    Figure Lengend Snippet:

    Article Snippet: Mouse monoclonal anti-human CD74 , eBioscience , Cat#11-0748-41; RRID: AB_2043845.

    Techniques: Recombinant, Coagulation, Antibody Labeling, Membrane, Plasmid Preparation, Polymer, Blocking Assay, Staining, Imaging, cDNA Synthesis, RNAscope, DNA Library Preparation, Software, Hybridization

    (a) Five melanoma cell lines were analyzed by Western blot analysis for CD74 expression. (b) Representative immunohistochemical staining intensities: score 0 (negative), 1 (weak), 2 (medium) and 3 (strong). Scale bar = 100 µm. (c) TMA analysis for CD74 expression during melanoma progression. CD74 staining was scored for the percentage (upper) and intensity (lower) of positive tumor cells. n indicates the number of tissue cores in each group. LN, lymph node. Both percentage and intensity score of each melanoma group were significantly higher than those for nevus group ( P < 0.0001).

    Journal: The Journal of investigative dermatology

    Article Title: Cell Surface CD74-MIF Interactions Drive Melanoma Survival in Response to Interferon-γ

    doi: 10.1038/jid.2015.204

    Figure Lengend Snippet: (a) Five melanoma cell lines were analyzed by Western blot analysis for CD74 expression. (b) Representative immunohistochemical staining intensities: score 0 (negative), 1 (weak), 2 (medium) and 3 (strong). Scale bar = 100 µm. (c) TMA analysis for CD74 expression during melanoma progression. CD74 staining was scored for the percentage (upper) and intensity (lower) of positive tumor cells. n indicates the number of tissue cores in each group. LN, lymph node. Both percentage and intensity score of each melanoma group were significantly higher than those for nevus group ( P < 0.0001).

    Article Snippet: Goat polyclonal anti-human actin (I-19; Santa Cruz, Carlsbad, CA), mouse monoclonal anti-human CD74 (PIN.1; Novus Biologicals, Littleton, CO), mouse monoclonal anti-human BCL-2 (124; GeneTex, San Antonio, TX), goat polyclonal anti-human MIF (R&D Systems, Minneapolis, MN) and rabbit monoclonal anti-human total AKT, phosphorylated AKT Ser473 (Cell Signaling Technology, Danvers, MA) antibodies were used as primary antibodies; horseradish peroxidase-labeled anti-mouse, anti-rabbit and anti-goat antibodies (1:1000; DAKO) were used as secondary antibodies.

    Techniques: Western Blot, Expressing, Immunohistochemical staining, Staining

    (a, b and c) Melanoma cell lines were treated with 0, 100 or 500 IU/ml IFN-γ for 48 hours and then assessed for CD74 expression. (a) Western blot analysis for total CD74 protein. Numbers above each band indicate relative CD74 expression levels. (b) Flow cytometric analysis for cell surface CD74. Histogram: black, isotype control; red, untreated cells stained with FITC-CD74 antibody; green, 100 IU/ml IFN-γ–treated cells with FITC-CD74 antibody; blue, 500 IU/ml IFN-γ–treated cells with FITC-CD74 antibody. (c) Immunofluorescence confocal microscopic analysis of cell surface CD74. Green (Alexa-488), CD74. Blue (DAPI), nucleus. A375 and SB2 cells showed slight expression under normal conditions (arrows). Scale bar = 30 µm. (d) Representative images of weak (left panels, score 1), medium (middle panels, score 2) and strong CD74 staining (right panels, score 3). Upper panel, Scale bar = 1 mm.; lower panel, Scale bar = 250 µm. Corresponding plasma IFN-γ levels are listed. (e) The association between CD74 staining at the tumor site and plasma IFN-γ levels in 55 patients with melanoma. Red bars, means. Error bars, standard deviations. ** P < 0.01, *** P < 0.001.

    Journal: The Journal of investigative dermatology

    Article Title: Cell Surface CD74-MIF Interactions Drive Melanoma Survival in Response to Interferon-γ

    doi: 10.1038/jid.2015.204

    Figure Lengend Snippet: (a, b and c) Melanoma cell lines were treated with 0, 100 or 500 IU/ml IFN-γ for 48 hours and then assessed for CD74 expression. (a) Western blot analysis for total CD74 protein. Numbers above each band indicate relative CD74 expression levels. (b) Flow cytometric analysis for cell surface CD74. Histogram: black, isotype control; red, untreated cells stained with FITC-CD74 antibody; green, 100 IU/ml IFN-γ–treated cells with FITC-CD74 antibody; blue, 500 IU/ml IFN-γ–treated cells with FITC-CD74 antibody. (c) Immunofluorescence confocal microscopic analysis of cell surface CD74. Green (Alexa-488), CD74. Blue (DAPI), nucleus. A375 and SB2 cells showed slight expression under normal conditions (arrows). Scale bar = 30 µm. (d) Representative images of weak (left panels, score 1), medium (middle panels, score 2) and strong CD74 staining (right panels, score 3). Upper panel, Scale bar = 1 mm.; lower panel, Scale bar = 250 µm. Corresponding plasma IFN-γ levels are listed. (e) The association between CD74 staining at the tumor site and plasma IFN-γ levels in 55 patients with melanoma. Red bars, means. Error bars, standard deviations. ** P < 0.01, *** P < 0.001.

    Article Snippet: Goat polyclonal anti-human actin (I-19; Santa Cruz, Carlsbad, CA), mouse monoclonal anti-human CD74 (PIN.1; Novus Biologicals, Littleton, CO), mouse monoclonal anti-human BCL-2 (124; GeneTex, San Antonio, TX), goat polyclonal anti-human MIF (R&D Systems, Minneapolis, MN) and rabbit monoclonal anti-human total AKT, phosphorylated AKT Ser473 (Cell Signaling Technology, Danvers, MA) antibodies were used as primary antibodies; horseradish peroxidase-labeled anti-mouse, anti-rabbit and anti-goat antibodies (1:1000; DAKO) were used as secondary antibodies.

    Techniques: Expressing, Western Blot, Control, Staining, Immunofluorescence, Clinical Proteomics

    (a and b) Cells were treated with ISO-1 for 48 hours and then analyzed by Western blot analysis for AKT Ser473 phosphorylation (a) and BCL-2 expression (b). Numbers above each band indicate the relative level of phosphorylated AKT Ser473 (pAKT) to total AKT (AKT) or BCL-2 to actin. (c, d and e) mRNA expression levels of BCL-2 (c), IL-6 (d) and IL-8 (e) in melanoma cells treated with ISO-1 for 24 hours were measured using qRT-PCR. (f) CD74 knockdown A375 cells were analyzed for mRNA expression of the indicated genes by qRT-PCR. shNT, non-target shRNA. * P < 0.05, ** P < 0.01.

    Journal: The Journal of investigative dermatology

    Article Title: Cell Surface CD74-MIF Interactions Drive Melanoma Survival in Response to Interferon-γ

    doi: 10.1038/jid.2015.204

    Figure Lengend Snippet: (a and b) Cells were treated with ISO-1 for 48 hours and then analyzed by Western blot analysis for AKT Ser473 phosphorylation (a) and BCL-2 expression (b). Numbers above each band indicate the relative level of phosphorylated AKT Ser473 (pAKT) to total AKT (AKT) or BCL-2 to actin. (c, d and e) mRNA expression levels of BCL-2 (c), IL-6 (d) and IL-8 (e) in melanoma cells treated with ISO-1 for 24 hours were measured using qRT-PCR. (f) CD74 knockdown A375 cells were analyzed for mRNA expression of the indicated genes by qRT-PCR. shNT, non-target shRNA. * P < 0.05, ** P < 0.01.

    Article Snippet: Goat polyclonal anti-human actin (I-19; Santa Cruz, Carlsbad, CA), mouse monoclonal anti-human CD74 (PIN.1; Novus Biologicals, Littleton, CO), mouse monoclonal anti-human BCL-2 (124; GeneTex, San Antonio, TX), goat polyclonal anti-human MIF (R&D Systems, Minneapolis, MN) and rabbit monoclonal anti-human total AKT, phosphorylated AKT Ser473 (Cell Signaling Technology, Danvers, MA) antibodies were used as primary antibodies; horseradish peroxidase-labeled anti-mouse, anti-rabbit and anti-goat antibodies (1:1000; DAKO) were used as secondary antibodies.

    Techniques: Western Blot, Phospho-proteomics, Expressing, Quantitative RT-PCR, Knockdown, shRNA

    Cell surface CD74-negative MeWo cells were subcutaneously injected into the flank of SCID Beige mice. Six days after cell injection, mice were randomly assigned to treatment groups (6 mice per group) and started on daily intraperitoneal treatment with ISO-1 (500 µg/mouse) and/or IFN-γ (1000 IU/mouse). ISO-1 and IFN-γ were administered for 21 days, and tumor growth was monitored. (a) Fold changes of tumor size. (b) Tumor weight on day 21. Error bars, standard deviations. * P < 0.05. (c) Representative mouse from each group (day 21). (d) Immunohistochemical staining for CD74 (upper) and MIF (lower) on day 21. Scale bar = 100 µm. ; AEC chromogen counterstained with hematoxylin.

    Journal: The Journal of investigative dermatology

    Article Title: Cell Surface CD74-MIF Interactions Drive Melanoma Survival in Response to Interferon-γ

    doi: 10.1038/jid.2015.204

    Figure Lengend Snippet: Cell surface CD74-negative MeWo cells were subcutaneously injected into the flank of SCID Beige mice. Six days after cell injection, mice were randomly assigned to treatment groups (6 mice per group) and started on daily intraperitoneal treatment with ISO-1 (500 µg/mouse) and/or IFN-γ (1000 IU/mouse). ISO-1 and IFN-γ were administered for 21 days, and tumor growth was monitored. (a) Fold changes of tumor size. (b) Tumor weight on day 21. Error bars, standard deviations. * P < 0.05. (c) Representative mouse from each group (day 21). (d) Immunohistochemical staining for CD74 (upper) and MIF (lower) on day 21. Scale bar = 100 µm. ; AEC chromogen counterstained with hematoxylin.

    Article Snippet: Goat polyclonal anti-human actin (I-19; Santa Cruz, Carlsbad, CA), mouse monoclonal anti-human CD74 (PIN.1; Novus Biologicals, Littleton, CO), mouse monoclonal anti-human BCL-2 (124; GeneTex, San Antonio, TX), goat polyclonal anti-human MIF (R&D Systems, Minneapolis, MN) and rabbit monoclonal anti-human total AKT, phosphorylated AKT Ser473 (Cell Signaling Technology, Danvers, MA) antibodies were used as primary antibodies; horseradish peroxidase-labeled anti-mouse, anti-rabbit and anti-goat antibodies (1:1000; DAKO) were used as secondary antibodies.

    Techniques: Injection, Immunohistochemical staining, Staining