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anti ck19  (Santa Cruz Biotechnology)


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    Structured Review

    Santa Cruz Biotechnology anti ck19
    Anti Ck19, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 95/100, based on 529 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/concentration+ck19/Cytokeratin+19+Antibody/pm28126348-317-101-103
    Average 95 stars, based on 529 article reviews
    anti ck19 - by Bioz Stars, 2026-10
    95/100 stars

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    Related Articles

    Co-Immunoprecipitation Assay:

    Article Title: Actomyosin forces trigger a conformational change in desmoplakin within desmosomes.
    Article Snippet: Phalloidin AF-488 (Invitrogen) and AF-568 (Invitrogen) was used for F-actin staining. .. The following antibodies were used for the Co-IP andwestern blot experiments: Anti-Keratin 19 (A53-B/A2), anti-plakophilin-3 (E-10), antiplakophilin-1 (10B2), anti-desmocolin-1 (A-4), anti-desmocolin-2/3 (7G6), anti-myosin-10 (A-3), and control mouse IgG (sc-2025) from Santa Cruz Biotechnology, anti-Keratin 8 (17514-1-AP), anti-Keratin 18 (10830-1-AP), anti-actin (66009-1-Ig), anti-desmoglein-2 (21880-1-AP), anti-plectin (29170-1-AP), anti-tubulin (66031-1-Ig), and antidesmoplakin (25318-1-AP) from Proteintech. .. The following secondary HRP conjugated antibodies were used: anti-mouse-HRP (A4416), and anti-rabbit-HRP (A6154) from Sigma Aldrich.

    Article Title: Actomyosin forces trigger a conformational change in desmoplakin within desmosomes
    Article Snippet: Phalloidin AF-488 (Invitrogen) and AF-568 (Invitrogen) was used for F-actin staining. .. The following antibodies were used for the Co-IP and western blot experiments: Anti-Keratin 19 (A53-B/A2), anti-plakophilin-3 (E-10), anti-plakophilin-1 (10B2), anti-desmocolin-1 (A-4), anti-desmocolin-2/3 (7G6), anti-myosin-10 (A-3), and control mouse IgG (sc-2025) from Santa Cruz Biotechnology, anti-Keratin 8 (17514-1-AP), anti-Keratin 18 (10830-1-AP), anti-actin (66009-1-Ig), anti-desmoglein-2 (21880-1-AP), anti-plectin (29170-1-AP), anti-tubulin (66031-1-Ig), and anti-desmoplakin (25318-1-AP) from Proteintech. .. The following secondary HRP conjugated antibodies were used: anti-mouse-HRP (A4416), and anti-rabbit-HRP (A6154) from Sigma Aldrich.

    Control:

    Article Title: Actomyosin forces trigger a conformational change in desmoplakin within desmosomes.
    Article Snippet: Phalloidin AF-488 (Invitrogen) and AF-568 (Invitrogen) was used for F-actin staining. .. The following antibodies were used for the Co-IP andwestern blot experiments: Anti-Keratin 19 (A53-B/A2), anti-plakophilin-3 (E-10), antiplakophilin-1 (10B2), anti-desmocolin-1 (A-4), anti-desmocolin-2/3 (7G6), anti-myosin-10 (A-3), and control mouse IgG (sc-2025) from Santa Cruz Biotechnology, anti-Keratin 8 (17514-1-AP), anti-Keratin 18 (10830-1-AP), anti-actin (66009-1-Ig), anti-desmoglein-2 (21880-1-AP), anti-plectin (29170-1-AP), anti-tubulin (66031-1-Ig), and antidesmoplakin (25318-1-AP) from Proteintech. .. The following secondary HRP conjugated antibodies were used: anti-mouse-HRP (A4416), and anti-rabbit-HRP (A6154) from Sigma Aldrich.

    Article Title: Actomyosin forces trigger a conformational change in desmoplakin within desmosomes
    Article Snippet: Phalloidin AF-488 (Invitrogen) and AF-568 (Invitrogen) was used for F-actin staining. .. The following antibodies were used for the Co-IP and western blot experiments: Anti-Keratin 19 (A53-B/A2), anti-plakophilin-3 (E-10), anti-plakophilin-1 (10B2), anti-desmocolin-1 (A-4), anti-desmocolin-2/3 (7G6), anti-myosin-10 (A-3), and control mouse IgG (sc-2025) from Santa Cruz Biotechnology, anti-Keratin 8 (17514-1-AP), anti-Keratin 18 (10830-1-AP), anti-actin (66009-1-Ig), anti-desmoglein-2 (21880-1-AP), anti-plectin (29170-1-AP), anti-tubulin (66031-1-Ig), and anti-desmoplakin (25318-1-AP) from Proteintech. .. The following secondary HRP conjugated antibodies were used: anti-mouse-HRP (A4416), and anti-rabbit-HRP (A6154) from Sigma Aldrich.

    Article Title: Compositions and methods for treating serpin B13 disorders
    Article Snippet: Isotype controls: TIB92 (10-3.6.2, mouse mAb, ATCC, 2.3 μg/mL), rabbit polyclonal IgG (cat. no. 02-6102, Invitrogen), PE rat IgG2a kappa (eBR2a) (cat. no. 12-4321-80, eBiosciense), Alexa Fluor 488 Rat IgG2a, kappa (cat. no. 400525, Biolegend) were diluted to the same concentrations. .. IF Microscopy: Anti-serpinB13 (clone B29, mouse mAb, 10 μg/mL), anti-serpinB13 (clone B34, mouse mAb, 10 μg/mL), TIB92 (10-3.6.2, mouse Isotype control, ATCC, 10 μg/mL), anti-cytokeratin 19 (clone B-1, mouse mAb, cat. no. Sc-374192, Santa Cruz, 1 μg/mL), anti-cytokeratin 17/19 (clone D4G2, rabbit mAb, cat. no. 12434S, Cell Signaling, 1:50), anti-cytokeratin 19-Alexa488 (clone EP1580Y, rabbit mAb, cat. no. ab192643, Abcam, 1:100), anti-Ngn3 (M-80, rabbit polyclonal antibody, cat. no. sc-25655, Santa Cruz, 2 μg/mL), anti-insulin (polyclonal guinea pig antibody, cat. no. A0564, DAKO, 2 μg/mL), anti-GFP (rabbit polyclonal antibody, cat. no. A21311, Life Technologies, 2 μg/mL). .. Secondary antibodies: Alexa Fluor 488 goat anti-mouse IgG (H+L) (cat. no. A11001, Invitrogen, 2 μg/mL), Alexa Fluor 488 goat anti-rabbit IgG (H+L), (cat no. A11034, Invitrogen, 2 g/mL), Alexa Fluor 568 goat anti-mouse IgG (H+L) (cat. no. A11031, Invitrogen, 5 μg/mL), Alexa Fluor 594 goat anti-guinea pig IgG (H+L) (cat. no. A11076, Invitrogen, 1 μg/mL), Alexa Fluor 568 goat anti-rabbit IgG (H+L), (cat. no. A11036, Invitrogen, 5 μg/mL), Alexa Fluor 647 goat anti-mouse IgG (H+L) (cat. no. A21235, Invitrogen, 5 μg/mL), and Alexa Fluor 647 goat anti-guinea pig IgG (H+L), (cat. no. A21450, Invitrogen, 5 μg/mL).

    Incubation:

    Article Title: The effect of stiffness on cell behavior and drug resistance in patient-derived breast cancer organoids.
    Article Snippet: Triple-negative breast cancer (TNBC) accounts for 10–15 % of breast cancer cases and is characterized by its high aggressiveness, influenced by extracellular matrix (ECM) proteins such as collagen IV and laminin.. Creating a cancer microenvironment that mimics these conditions could enhance the clinical relevance of breast cancer models for patient-specific treatments.. In this study, a Segmented Organoids with Agile Reassembly (SOAR) printing was established by compartmentalizing TNBC spheroids with ECM proteins (Matrigel®) to simulate varying levels of aggressiveness.

    Article Title: Long-term SiNPs exposure induced genetic instability and malignant transformation via SQSTM1/p62-mediated autophagy dysfunction in lungs
    Article Snippet: .. Sections were incubated overnight at 4 °C with primary antibodies against γ-H2AX (Abcam, ab81299), SQSTM1 (Abcam, ab109012), LC3B (CST, 12741), and CK-19 (Santa Cruz, sc-376126). ..

    Article Title: Key genes and histopathological alterations in hepatic fibrogenesis after segmental cholestasis in weaned rats.
    Article Snippet: BACKGROUND: The selective bile duct ligation (sBDL) model has been proposed to study fibrosis progression in segmental cholestasis, replicating histopathological features of biliary obstruction in both liver parenchyma with biliary obstruction (BO) and without biliary obstruction (WBO).. However, the molecular mechanisms driving fibrogenesis in WBO parenchyma remain unclear.. This study aimed to characterize gene expression and histological alterations in fibrogenesis using the sBDL model. METHODS: Liver samples were collected at 1, 4, and 8 weeks post-surgery from both BO and WBO parenchyma, undergoing histological, immunohistochemical, biochemical, and molecular analyses.

    Flow Cytometry:

    Article Title: Compositions and methods for treating serpin B13 disorders
    Article Snippet: Secondary antibodies: HRP-conjugated anti-sheep IgG (polyclonal donkey IgG, cat. no. HAF016, Bio-Techne, 1:1000), HRP-conjugated anti-rabbit IgG (polygoclonal goat IgG, cat. no. A27036, Invitrogen, 0.1 μg/mL) and HRP-conjugated anti-mouse IgG (rabbit polygoclonal, cat. no. A27025, Invitrogen, 0.1 g/mL). .. Flow Cytometry: Anti-serpinB13 (clone B29, mouse mAb, 2.3 μg/mL), anti-cytokeratin19 (clone B-1, mouse mAb, cat. no. sc-374192, Santa Cruz, 1 μg/mL), anti-Ngn3 (M-80, rabbit polyclonal, cat. no. Sc25655, Santa Cruz, 1 μg/mL), anti-Notch1-PE (clone 22E5, rat mAb, cat. no. 12-5765-82, eBioscience, 1 μg/mL), anti-activated Notch1 (Val1744, rabbit polyclonal whole antiserum, cat. no. ab8925, Abcam, 1:800), anti-EpCAM-Alexa488 (clone G8.8, rat IgG, cat. no. 118210, BioLegend, 0.625 μg/mL), anti-insulin-Alexa647 (clone T56-706, mouse mAb, cat. no. 565689, BD Biosciences, 1 μg/mL), anti-glucagon-PE (clone U16-850, mouse mAb cat. no. 565860, BD Biosciences, 1:400). .. Secondary antibodies: Alexa Fluor 488 goat anti-mouse IgG (H+L) (cat. no. A11001, Invitrogen, 2 μg/mL) and Alexa Fluor 568 goat anti-rabbit IgG (H+L) (cat. no. A11036, Invitrogen, 2 μg/mL).

    Western Blot:

    Article Title: Actomyosin forces trigger a conformational change in desmoplakin within desmosomes
    Article Snippet: Phalloidin AF-488 (Invitrogen) and AF-568 (Invitrogen) was used for F-actin staining. .. The following antibodies were used for the Co-IP and western blot experiments: Anti-Keratin 19 (A53-B/A2), anti-plakophilin-3 (E-10), anti-plakophilin-1 (10B2), anti-desmocolin-1 (A-4), anti-desmocolin-2/3 (7G6), anti-myosin-10 (A-3), and control mouse IgG (sc-2025) from Santa Cruz Biotechnology, anti-Keratin 8 (17514-1-AP), anti-Keratin 18 (10830-1-AP), anti-actin (66009-1-Ig), anti-desmoglein-2 (21880-1-AP), anti-plectin (29170-1-AP), anti-tubulin (66031-1-Ig), and anti-desmoplakin (25318-1-AP) from Proteintech. .. The following secondary HRP conjugated antibodies were used: anti-mouse-HRP (A4416), and anti-rabbit-HRP (A6154) from Sigma Aldrich.

    Microscopy:

    Article Title: Compositions and methods for treating serpin B13 disorders
    Article Snippet: Isotype controls: TIB92 (10-3.6.2, mouse mAb, ATCC, 2.3 μg/mL), rabbit polyclonal IgG (cat. no. 02-6102, Invitrogen), PE rat IgG2a kappa (eBR2a) (cat. no. 12-4321-80, eBiosciense), Alexa Fluor 488 Rat IgG2a, kappa (cat. no. 400525, Biolegend) were diluted to the same concentrations. .. IF Microscopy: Anti-serpinB13 (clone B29, mouse mAb, 10 μg/mL), anti-serpinB13 (clone B34, mouse mAb, 10 μg/mL), TIB92 (10-3.6.2, mouse Isotype control, ATCC, 10 μg/mL), anti-cytokeratin 19 (clone B-1, mouse mAb, cat. no. Sc-374192, Santa Cruz, 1 μg/mL), anti-cytokeratin 17/19 (clone D4G2, rabbit mAb, cat. no. 12434S, Cell Signaling, 1:50), anti-cytokeratin 19-Alexa488 (clone EP1580Y, rabbit mAb, cat. no. ab192643, Abcam, 1:100), anti-Ngn3 (M-80, rabbit polyclonal antibody, cat. no. sc-25655, Santa Cruz, 2 μg/mL), anti-insulin (polyclonal guinea pig antibody, cat. no. A0564, DAKO, 2 μg/mL), anti-GFP (rabbit polyclonal antibody, cat. no. A21311, Life Technologies, 2 μg/mL). .. Secondary antibodies: Alexa Fluor 488 goat anti-mouse IgG (H+L) (cat. no. A11001, Invitrogen, 2 μg/mL), Alexa Fluor 488 goat anti-rabbit IgG (H+L), (cat no. A11034, Invitrogen, 2 g/mL), Alexa Fluor 568 goat anti-mouse IgG (H+L) (cat. no. A11031, Invitrogen, 5 μg/mL), Alexa Fluor 594 goat anti-guinea pig IgG (H+L) (cat. no. A11076, Invitrogen, 1 μg/mL), Alexa Fluor 568 goat anti-rabbit IgG (H+L), (cat. no. A11036, Invitrogen, 5 μg/mL), Alexa Fluor 647 goat anti-mouse IgG (H+L) (cat. no. A21235, Invitrogen, 5 μg/mL), and Alexa Fluor 647 goat anti-guinea pig IgG (H+L), (cat. no. A21450, Invitrogen, 5 μg/mL).



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    (A) Graphic representation of alleles altered in Ptf1aCreERTM mice. Triangles represent LoxP sites. Arrows indicate genetic loci. Grey boxes indicate exons. (B) Bioluminescence imaging demonstrating tamoxifen (Tam) induced Ptf1aCreERTM activity; images are representative of 3 separate experiments involving 2 mice per group. Radiance unit is photons/second/cm2/steradian (C) Multiplexed IF micrographs indicating expression of lineage marker (mCherry) only in acinar cells. Micrographs depict acinar cells (upper panels) and pancreatic ductal cells (lower panels). Channels are DAPI (blue), mCherry (white), Cpa1 (green), and <t>CK19</t> (red). Micrographs are representative of n = 2 Ptf1aCre-ERTM positive mice. Magnification 63X, scale bars represent 100 μm.
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    Image Search Results


    Journal: Medicina Oral, Patología Oral y Cirugía Bucal

    Article Title: Clinical and Immunohistochemical epithelial profile of non-healing chronic traumatic ulcers

    doi: 10.4317/medoral.23729

    Figure Lengend Snippet: Immunolabelling for Ck19 and Ki67.

    Article Snippet: The following primary antibodies were used: concentrated rabbit anti-human Ki67 (CellMarque, SIGMA ALDRICH, USA), and concentrated monoclonal mouse anti-human Ck19 (CellMarque, SIGMA ALDRICH, USA).

    Techniques:

    Immunolabeling of Ck19 and Ki67 for Non-healing CTU and OSCC group. The black arrow indicates positive immunolabeling of nuclei and white asterisk indicates positive immunolabeling of cytoplasm. The immunostaining was considered negative when observed 0–5% positive cells. The analysis of the samples was performed by a single-blind examiner. The positive immunostaining was counted for each group and expressed. Stacked bars represent the percentage of immunolabelled samples for each biomarker used according to their score (negative low-medium-high). The sections were analyzed by optical microscopy at a magnification of 20X and 400 X. p <0.05 indicates statistical significance. 2A- OSCC of the lateral tongue diagnosed at an early stage. 2B- Islet cells with Ck19 positive cytoplasm. OSCC group. 2C-High index of Ki67, OSCC group. 2D- Non healing Chronic Traumatic Ulcer located on the lateral tongue associated with sharp cups and parafunctional habits (tongue interposition). The ulcer did not heal after 15 days of the CMI source control. 2E Niches of Ck19 positive cells with a suprabasal pattern found within the hyperplastic epithelium of the edges of the NHCTU. 2F Suprabasal expression of Ki67, NHCTU group. Nuclear alterations were evidenced with the Ki67 immunostaining.

    Journal: Medicina Oral, Patología Oral y Cirugía Bucal

    Article Title: Clinical and Immunohistochemical epithelial profile of non-healing chronic traumatic ulcers

    doi: 10.4317/medoral.23729

    Figure Lengend Snippet: Immunolabeling of Ck19 and Ki67 for Non-healing CTU and OSCC group. The black arrow indicates positive immunolabeling of nuclei and white asterisk indicates positive immunolabeling of cytoplasm. The immunostaining was considered negative when observed 0–5% positive cells. The analysis of the samples was performed by a single-blind examiner. The positive immunostaining was counted for each group and expressed. Stacked bars represent the percentage of immunolabelled samples for each biomarker used according to their score (negative low-medium-high). The sections were analyzed by optical microscopy at a magnification of 20X and 400 X. p <0.05 indicates statistical significance. 2A- OSCC of the lateral tongue diagnosed at an early stage. 2B- Islet cells with Ck19 positive cytoplasm. OSCC group. 2C-High index of Ki67, OSCC group. 2D- Non healing Chronic Traumatic Ulcer located on the lateral tongue associated with sharp cups and parafunctional habits (tongue interposition). The ulcer did not heal after 15 days of the CMI source control. 2E Niches of Ck19 positive cells with a suprabasal pattern found within the hyperplastic epithelium of the edges of the NHCTU. 2F Suprabasal expression of Ki67, NHCTU group. Nuclear alterations were evidenced with the Ki67 immunostaining.

    Article Snippet: The following primary antibodies were used: concentrated rabbit anti-human Ki67 (CellMarque, SIGMA ALDRICH, USA), and concentrated monoclonal mouse anti-human Ck19 (CellMarque, SIGMA ALDRICH, USA).

    Techniques: Immunolabeling, Immunostaining, Biomarker Discovery, Microscopy, Control, Expressing

    The epithelium homeostasis of the oral mucosa involves a balance between cell differentiation and cell proliferation. CMI could generate a stimulus in the increase of immature Ck19 cells (OKSC) and a continuous presence of these cells, known for their genomic instability, in the suprabasal layers of the oral epithelium. These cells, considered as a target of different carcinogens, are also involved in the self-renewal of the epithelium and healing events. An epithelial environment of hyperproliferation and cellular immaturity is generated chronically next to a wound with no tendency to cure. CMI could act as a promoter of oral carcinogenesis through these supposed mechanisms.

    Journal: Medicina Oral, Patología Oral y Cirugía Bucal

    Article Title: Clinical and Immunohistochemical epithelial profile of non-healing chronic traumatic ulcers

    doi: 10.4317/medoral.23729

    Figure Lengend Snippet: The epithelium homeostasis of the oral mucosa involves a balance between cell differentiation and cell proliferation. CMI could generate a stimulus in the increase of immature Ck19 cells (OKSC) and a continuous presence of these cells, known for their genomic instability, in the suprabasal layers of the oral epithelium. These cells, considered as a target of different carcinogens, are also involved in the self-renewal of the epithelium and healing events. An epithelial environment of hyperproliferation and cellular immaturity is generated chronically next to a wound with no tendency to cure. CMI could act as a promoter of oral carcinogenesis through these supposed mechanisms.

    Article Snippet: The following primary antibodies were used: concentrated rabbit anti-human Ki67 (CellMarque, SIGMA ALDRICH, USA), and concentrated monoclonal mouse anti-human Ck19 (CellMarque, SIGMA ALDRICH, USA).

    Techniques: Cell Differentiation, Generated

    (A) Graphic representation of alleles altered in Ptf1aCreERTM mice. Triangles represent LoxP sites. Arrows indicate genetic loci. Grey boxes indicate exons. (B) Bioluminescence imaging demonstrating tamoxifen (Tam) induced Ptf1aCreERTM activity; images are representative of 3 separate experiments involving 2 mice per group. Radiance unit is photons/second/cm2/steradian (C) Multiplexed IF micrographs indicating expression of lineage marker (mCherry) only in acinar cells. Micrographs depict acinar cells (upper panels) and pancreatic ductal cells (lower panels). Channels are DAPI (blue), mCherry (white), Cpa1 (green), and CK19 (red). Micrographs are representative of n = 2 Ptf1aCre-ERTM positive mice. Magnification 63X, scale bars represent 100 μm.

    Journal: PLoS ONE

    Article Title: Desmoplasia and oncogene driven acinar-to-ductal metaplasia are concurrent events during acinar cell-derived pancreatic cancer initiation in young adult mice

    doi: 10.1371/journal.pone.0221810

    Figure Lengend Snippet: (A) Graphic representation of alleles altered in Ptf1aCreERTM mice. Triangles represent LoxP sites. Arrows indicate genetic loci. Grey boxes indicate exons. (B) Bioluminescence imaging demonstrating tamoxifen (Tam) induced Ptf1aCreERTM activity; images are representative of 3 separate experiments involving 2 mice per group. Radiance unit is photons/second/cm2/steradian (C) Multiplexed IF micrographs indicating expression of lineage marker (mCherry) only in acinar cells. Micrographs depict acinar cells (upper panels) and pancreatic ductal cells (lower panels). Channels are DAPI (blue), mCherry (white), Cpa1 (green), and CK19 (red). Micrographs are representative of n = 2 Ptf1aCre-ERTM positive mice. Magnification 63X, scale bars represent 100 μm.

    Article Snippet: Primary antibodies were diluted in PBS-T and used at the following concentration CK19 1/20 (DSHB Cat# TROMA-III, RRID:AB_2133570), Cpa1 1/1000 (R&D Systems Cat# AF2765, RRID:AB_2085841), Sox9 1/5000 (Millipore Cat# AB5535, RRID:AB_2239761), αSMA 1/200 (Thermo Fisher Scientific Cat# 14-9760-80, RRID:AB_2572995), mCherry 1/50 (Rockland Cat# 600-401-379, RRID:AB_2209751).

    Techniques: Imaging, Activity Assay, Expressing, Marker

    (A) Graphic representation of alleles altered in KPT mice. Triangles represent LoxP sites. Arrows indicate genetic loci. Grey boxes indicate exons. Asterisks indicate point mutations. (B) Kaplan-Meier survival analysis of KPT and KP mice post tamoxifen injection. Median survival for KPT mice is 219 day post tamoxifen injection. P < 0.0001 by means of the log-rank test. Representative micrographs depicting H&E stained sections of PDAC (C) and liver metastasis (D) in KPT mice. (E) Representative micrographs of Trichrome stained tissue sections of PDAC in KPT mice ( n = 5). (F) Representative micrographs depicting histochemical visualization of HA in KPT tumors, HA is stained brown, nuclei are counterstained blue ( n = 5). Magnification of brightfield images is 20x. (G) Multiplexed IF staining of KPT tumors for detection of nuclei (DAPI, blue), Sox9 (white) and CK19 (red). n = 3 KPT tumors, magnification 40x. (H) Multiplexed IF staining of KPT tumors for detection of nuclei (DAPI, blue), mCherry (white) and CK19 (red) n = 2 KPT tumors, magnification 40x. (I) Multiplexed IF staining of KPT tumors for detection of nuclei (DAPI, blue), αSMA (white) and CK19 (red) n = 3 KPT tumors, magnification 63x. All scale bars represent 100 μm.

    Journal: PLoS ONE

    Article Title: Desmoplasia and oncogene driven acinar-to-ductal metaplasia are concurrent events during acinar cell-derived pancreatic cancer initiation in young adult mice

    doi: 10.1371/journal.pone.0221810

    Figure Lengend Snippet: (A) Graphic representation of alleles altered in KPT mice. Triangles represent LoxP sites. Arrows indicate genetic loci. Grey boxes indicate exons. Asterisks indicate point mutations. (B) Kaplan-Meier survival analysis of KPT and KP mice post tamoxifen injection. Median survival for KPT mice is 219 day post tamoxifen injection. P < 0.0001 by means of the log-rank test. Representative micrographs depicting H&E stained sections of PDAC (C) and liver metastasis (D) in KPT mice. (E) Representative micrographs of Trichrome stained tissue sections of PDAC in KPT mice ( n = 5). (F) Representative micrographs depicting histochemical visualization of HA in KPT tumors, HA is stained brown, nuclei are counterstained blue ( n = 5). Magnification of brightfield images is 20x. (G) Multiplexed IF staining of KPT tumors for detection of nuclei (DAPI, blue), Sox9 (white) and CK19 (red). n = 3 KPT tumors, magnification 40x. (H) Multiplexed IF staining of KPT tumors for detection of nuclei (DAPI, blue), mCherry (white) and CK19 (red) n = 2 KPT tumors, magnification 40x. (I) Multiplexed IF staining of KPT tumors for detection of nuclei (DAPI, blue), αSMA (white) and CK19 (red) n = 3 KPT tumors, magnification 63x. All scale bars represent 100 μm.

    Article Snippet: Primary antibodies were diluted in PBS-T and used at the following concentration CK19 1/20 (DSHB Cat# TROMA-III, RRID:AB_2133570), Cpa1 1/1000 (R&D Systems Cat# AF2765, RRID:AB_2085841), Sox9 1/5000 (Millipore Cat# AB5535, RRID:AB_2239761), αSMA 1/200 (Thermo Fisher Scientific Cat# 14-9760-80, RRID:AB_2572995), mCherry 1/50 (Rockland Cat# 600-401-379, RRID:AB_2209751).

    Techniques: Injection, Staining

    (A-B) Representative micrographs depicting H&E stained sections of the pancreata of KP (A) and KPT (B) mice 3 months post tamoxifen treatment. Black arrows indicate regions of acinar to ductal metaplasia, red arrow indicates PanIN lesion. ( n = 6 KPT mice, and n = 7 KP mice), magnification 20x. (C) Representative multiplexed IF micrographs of KPT pancreata 2 months post tamoxifen treatment, for detection of nuclei (DAPI, blue), Cpa1 (green), Sox9 (white) and CK19 (red). Images depict acinar tissue with normal histology (upper panels), an isolated region of early acinar to ductal metaplasia (middle panels) and a PanIN lesion (lower panels), n = 6 KPT mice, magnification is 63X. (D) Representative micrographs of alcian blue and eosin (AB/E) stained sections of the pancreata of KPT and KP littermate control mice 1M, 2M, and 3M post tamoxifen injection, magnification is 20X. (E) Whole slide quantification of alcian blue positive area relative to eosin positive total pancreatic area, data points represent 1 AB/E stained slide per mouse, n = 7 for KP mice, n = 4 for 1M KPT mice, n = 6 for 2M KPT, n = 6 for 3M KPT. Bars depicts mean with error bars representing standard error of the mean (SEM). Significance was determined by student’s t test. *, P < 0.05; ** , P < 0.01; *** , P < 0.001. All scale bars represent 100 μm.

    Journal: PLoS ONE

    Article Title: Desmoplasia and oncogene driven acinar-to-ductal metaplasia are concurrent events during acinar cell-derived pancreatic cancer initiation in young adult mice

    doi: 10.1371/journal.pone.0221810

    Figure Lengend Snippet: (A-B) Representative micrographs depicting H&E stained sections of the pancreata of KP (A) and KPT (B) mice 3 months post tamoxifen treatment. Black arrows indicate regions of acinar to ductal metaplasia, red arrow indicates PanIN lesion. ( n = 6 KPT mice, and n = 7 KP mice), magnification 20x. (C) Representative multiplexed IF micrographs of KPT pancreata 2 months post tamoxifen treatment, for detection of nuclei (DAPI, blue), Cpa1 (green), Sox9 (white) and CK19 (red). Images depict acinar tissue with normal histology (upper panels), an isolated region of early acinar to ductal metaplasia (middle panels) and a PanIN lesion (lower panels), n = 6 KPT mice, magnification is 63X. (D) Representative micrographs of alcian blue and eosin (AB/E) stained sections of the pancreata of KPT and KP littermate control mice 1M, 2M, and 3M post tamoxifen injection, magnification is 20X. (E) Whole slide quantification of alcian blue positive area relative to eosin positive total pancreatic area, data points represent 1 AB/E stained slide per mouse, n = 7 for KP mice, n = 4 for 1M KPT mice, n = 6 for 2M KPT, n = 6 for 3M KPT. Bars depicts mean with error bars representing standard error of the mean (SEM). Significance was determined by student’s t test. *, P < 0.05; ** , P < 0.01; *** , P < 0.001. All scale bars represent 100 μm.

    Article Snippet: Primary antibodies were diluted in PBS-T and used at the following concentration CK19 1/20 (DSHB Cat# TROMA-III, RRID:AB_2133570), Cpa1 1/1000 (R&D Systems Cat# AF2765, RRID:AB_2085841), Sox9 1/5000 (Millipore Cat# AB5535, RRID:AB_2239761), αSMA 1/200 (Thermo Fisher Scientific Cat# 14-9760-80, RRID:AB_2572995), mCherry 1/50 (Rockland Cat# 600-401-379, RRID:AB_2209751).

    Techniques: Staining, Isolation, Control, Injection

    (A) Histochemical visualization of HA (brown) counterstained with hematoxylin (blue). Representative micrographs depict the pancreata of KPT and KP mice 1 month, 2 months, and 3 months post-tamoxifen injection. Images of KPT pancreata depict dysplastic regions containing ADM and/or PanIN. Masson’s trichrome staining depicting collagen deposition (blue) in serial sections from the same KPT pancreata as shown in (A) , n = 7 for KP mice, n = 4 for 1M KPT mice, n = 6 for 2M KPT, n = 6 for 3M KPT. Magnification of bright field images is 20X. (B) Representative multiplexed IF micrographs for detection of nuclei (DAPI, blue), αSMA (white), CK19 (red), and Cpa1 (green). Rows depict healthy acinar tissue (acinar), a region of ADM and PanIN lesions. Micrographs are representative of the pancreata of KPT mice 2 months post tamoxifen treatment ( n = 3), magnification is 63X. All scale bars represent 100 μm.We sought to identify the source of HA in regions of ADM, via the detection of activated PSC. Increased activation and proliferation of PSC are directly involved in the development of PDAC associated fibrosis via the deposition of ECM components including HA and collagen in the tumor microenvironment . Multiplexed IF staining against αSMA, CK19 and Cpa1 revealed the presence of activated PSC in proximity to PanIN lesions and infiltrating regions of ADM in KPT mice. Activated αSMA-positive PSCs were not observed in regions of Cpa1 + /CK19 - healthy acinar tissue ( Fig 5B , top panel). Activated PSC were observed infiltrating regions of ADM containing Cpa1 + /CK19 + cells undergoing ADM ( Fig 5B , middle panel), and CK19 + /Cpa1 - PanIN lesions were surrounded by activated PSC ( Fig 5B , bottom panel). These observations indicate that PSC recruitment and/or activation occur during ADM and that neoplasia-associated HA production and PSC activation precede collagen deposition, which is associated with PanIN lesions. To our knowledge, the role of desmoplasia in general, or HA deposition specifically, in the initiation of PDAC has not been investigated in detail, due to the paucity of animal models. While PSC activation in regions of ADM is associated with HA deposition, additional experiments are necessary to definitively identify the specific cellular origin of HA in regions of ADM. These results establish the KPT model as an ideal model to study the role of the desmoplastic response in PDAC development from a defined population of adult cells.

    Journal: PLoS ONE

    Article Title: Desmoplasia and oncogene driven acinar-to-ductal metaplasia are concurrent events during acinar cell-derived pancreatic cancer initiation in young adult mice

    doi: 10.1371/journal.pone.0221810

    Figure Lengend Snippet: (A) Histochemical visualization of HA (brown) counterstained with hematoxylin (blue). Representative micrographs depict the pancreata of KPT and KP mice 1 month, 2 months, and 3 months post-tamoxifen injection. Images of KPT pancreata depict dysplastic regions containing ADM and/or PanIN. Masson’s trichrome staining depicting collagen deposition (blue) in serial sections from the same KPT pancreata as shown in (A) , n = 7 for KP mice, n = 4 for 1M KPT mice, n = 6 for 2M KPT, n = 6 for 3M KPT. Magnification of bright field images is 20X. (B) Representative multiplexed IF micrographs for detection of nuclei (DAPI, blue), αSMA (white), CK19 (red), and Cpa1 (green). Rows depict healthy acinar tissue (acinar), a region of ADM and PanIN lesions. Micrographs are representative of the pancreata of KPT mice 2 months post tamoxifen treatment ( n = 3), magnification is 63X. All scale bars represent 100 μm.We sought to identify the source of HA in regions of ADM, via the detection of activated PSC. Increased activation and proliferation of PSC are directly involved in the development of PDAC associated fibrosis via the deposition of ECM components including HA and collagen in the tumor microenvironment . Multiplexed IF staining against αSMA, CK19 and Cpa1 revealed the presence of activated PSC in proximity to PanIN lesions and infiltrating regions of ADM in KPT mice. Activated αSMA-positive PSCs were not observed in regions of Cpa1 + /CK19 - healthy acinar tissue ( Fig 5B , top panel). Activated PSC were observed infiltrating regions of ADM containing Cpa1 + /CK19 + cells undergoing ADM ( Fig 5B , middle panel), and CK19 + /Cpa1 - PanIN lesions were surrounded by activated PSC ( Fig 5B , bottom panel). These observations indicate that PSC recruitment and/or activation occur during ADM and that neoplasia-associated HA production and PSC activation precede collagen deposition, which is associated with PanIN lesions. To our knowledge, the role of desmoplasia in general, or HA deposition specifically, in the initiation of PDAC has not been investigated in detail, due to the paucity of animal models. While PSC activation in regions of ADM is associated with HA deposition, additional experiments are necessary to definitively identify the specific cellular origin of HA in regions of ADM. These results establish the KPT model as an ideal model to study the role of the desmoplastic response in PDAC development from a defined population of adult cells.

    Article Snippet: Primary antibodies were diluted in PBS-T and used at the following concentration CK19 1/20 (DSHB Cat# TROMA-III, RRID:AB_2133570), Cpa1 1/1000 (R&D Systems Cat# AF2765, RRID:AB_2085841), Sox9 1/5000 (Millipore Cat# AB5535, RRID:AB_2239761), αSMA 1/200 (Thermo Fisher Scientific Cat# 14-9760-80, RRID:AB_2572995), mCherry 1/50 (Rockland Cat# 600-401-379, RRID:AB_2209751).

    Techniques: Injection, Staining, Activation Assay