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codex staining kit  (Akoya Biosciences)


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    Akoya Biosciences codex staining kit
    KITL regulates PSC state and pancreas tissue homeostasis. A, Volcano plot of all upregulated, nonsignificant, and downregulated differentially expressed genes (DEG) as defined by the Wald test ( P .adj < 0.05 and log 2 FC > 1) from Kitl overexpression ( Kitl OE) ImPSC-1 bulk RNA-seq dataset with representative gene labels included. Data represent three biological repeats. B, Gene ontology (GO) analysis of upregulated and downregulated genes in immortalized PSCs (ImPSC-1) overexpressing Kitl . Top 10 enrichment categories ranked by adjusted P values plotted in each direction. C, qRT-PCR analysis of indicated DEGs in ImPSC-1 expressing stable Kitl knockdown (sh Kitl ) or overexpression ( Kitl OE). PLKO vector (shCtrl) and fluorescence overexpression ( Egfp OE) serves as the respective control. D, Representative images of <t>CODEX</t> <t>staining</t> (left) and quantification (right) for GFP (green) and CD31 (red) in normal pancreas from Fabp4-Cre or Kitl fl/fl ;Fabp4-Cre mouse model ( n = 3–4 mice per arm). Scale bar, 100 μm. E, Representative images of CODEX composite staining (left) and quantification (right) for CD45 (white) in normal pancreas from Fabp4-Cre or Kitl fl/fl ;Fabp4-Cre mouse model ( n = 4 mice per arm). Scale bar, 50 μm. F, Representative images of IHC staining for GFP (green), c-KIT receptor (KIT, red), and VIM (magenta) in healthy murine pancreas from Rosa26 mTmG/+ ;Fabp4-Cre mice. Scale bar, 10 μm. G, Representative images of IHC staining for CD31 (green) and c-KIT receptor (KIT, red) in healthy murine pancreas. Scale bar, 10 μm. H, Representative images of IHC staining for panCK (white) and c-KIT receptor (KIT, red) in healthy murine pancreas. Scale bar, 10 μm. I, Representative hematoxylin and eosin images between caerulein-treated Fabp4-Cre and Kitl fl/fl ;Fabp4-Cre mice. Scale bar, 100 μm. J, Representative images of IHC staining (left) for panCK (green) and amylase (red) between caerulein-treated Fabp4-Cre and Kitl fl/fl ;Fabp4-Cre mice. Scale bar, 10 μm. PanCK quantification on right ( n = 3 mice per arm). K, Representative IHC images (left) and quantification (right) for CD45 staining between caerulein-treated Fabp4-Cre and Kitl fl/fl ;Fabp4-Cre mice. Scale bar, 100 μm. L, Representative images (left) and quantification (right) for CD68 staining in caerulein-treated Fabp4-Cre and Kitl fl/fl ;Fabp4-Cre mouse pancreas. The same co-stained tissue section is shown in K and L . Scale bar, 100 μm. For comparisons of two groups, an unpaired t test was used. For comparisons of more than two groups with one independent variable, the one-way ANOVA test was used. Data are represented as mean ± SEM; *, P ≤ 0.05; **, P ≤ 0.01; ***, P ≤ 0.001; ****, P ≤ 0.0001; FC, fold changes; ns, not significant; panc area, pancreas.
    Codex Staining Kit, supplied by Akoya Biosciences, used in various techniques. Bioz Stars score: 99/100, based on 12644 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/codex+staining+kit/PhenoCycler-Fusion+2%2E0/pmc12046321-314-27-30
    Average 99 stars, based on 12644 article reviews
    codex staining kit - by Bioz Stars, 2026-11
    99/100 stars

    Images

    1) Product Images from "Stromal KITL/SCF Maintains Pancreas Tissue Homeostasis and Restrains Tumor Progression"

    Article Title: Stromal KITL/SCF Maintains Pancreas Tissue Homeostasis and Restrains Tumor Progression

    Journal: Cancer Discovery

    doi: 10.1158/2159-8290.CD-24-1079

    KITL regulates PSC state and pancreas tissue homeostasis. A, Volcano plot of all upregulated, nonsignificant, and downregulated differentially expressed genes (DEG) as defined by the Wald test ( P .adj < 0.05 and log 2 FC > 1) from Kitl overexpression ( Kitl OE) ImPSC-1 bulk RNA-seq dataset with representative gene labels included. Data represent three biological repeats. B, Gene ontology (GO) analysis of upregulated and downregulated genes in immortalized PSCs (ImPSC-1) overexpressing Kitl . Top 10 enrichment categories ranked by adjusted P values plotted in each direction. C, qRT-PCR analysis of indicated DEGs in ImPSC-1 expressing stable Kitl knockdown (sh Kitl ) or overexpression ( Kitl OE). PLKO vector (shCtrl) and fluorescence overexpression ( Egfp OE) serves as the respective control. D, Representative images of CODEX staining (left) and quantification (right) for GFP (green) and CD31 (red) in normal pancreas from Fabp4-Cre or Kitl fl/fl ;Fabp4-Cre mouse model ( n = 3–4 mice per arm). Scale bar, 100 μm. E, Representative images of CODEX composite staining (left) and quantification (right) for CD45 (white) in normal pancreas from Fabp4-Cre or Kitl fl/fl ;Fabp4-Cre mouse model ( n = 4 mice per arm). Scale bar, 50 μm. F, Representative images of IHC staining for GFP (green), c-KIT receptor (KIT, red), and VIM (magenta) in healthy murine pancreas from Rosa26 mTmG/+ ;Fabp4-Cre mice. Scale bar, 10 μm. G, Representative images of IHC staining for CD31 (green) and c-KIT receptor (KIT, red) in healthy murine pancreas. Scale bar, 10 μm. H, Representative images of IHC staining for panCK (white) and c-KIT receptor (KIT, red) in healthy murine pancreas. Scale bar, 10 μm. I, Representative hematoxylin and eosin images between caerulein-treated Fabp4-Cre and Kitl fl/fl ;Fabp4-Cre mice. Scale bar, 100 μm. J, Representative images of IHC staining (left) for panCK (green) and amylase (red) between caerulein-treated Fabp4-Cre and Kitl fl/fl ;Fabp4-Cre mice. Scale bar, 10 μm. PanCK quantification on right ( n = 3 mice per arm). K, Representative IHC images (left) and quantification (right) for CD45 staining between caerulein-treated Fabp4-Cre and Kitl fl/fl ;Fabp4-Cre mice. Scale bar, 100 μm. L, Representative images (left) and quantification (right) for CD68 staining in caerulein-treated Fabp4-Cre and Kitl fl/fl ;Fabp4-Cre mouse pancreas. The same co-stained tissue section is shown in K and L . Scale bar, 100 μm. For comparisons of two groups, an unpaired t test was used. For comparisons of more than two groups with one independent variable, the one-way ANOVA test was used. Data are represented as mean ± SEM; *, P ≤ 0.05; **, P ≤ 0.01; ***, P ≤ 0.001; ****, P ≤ 0.0001; FC, fold changes; ns, not significant; panc area, pancreas.
    Figure Legend Snippet: KITL regulates PSC state and pancreas tissue homeostasis. A, Volcano plot of all upregulated, nonsignificant, and downregulated differentially expressed genes (DEG) as defined by the Wald test ( P .adj < 0.05 and log 2 FC > 1) from Kitl overexpression ( Kitl OE) ImPSC-1 bulk RNA-seq dataset with representative gene labels included. Data represent three biological repeats. B, Gene ontology (GO) analysis of upregulated and downregulated genes in immortalized PSCs (ImPSC-1) overexpressing Kitl . Top 10 enrichment categories ranked by adjusted P values plotted in each direction. C, qRT-PCR analysis of indicated DEGs in ImPSC-1 expressing stable Kitl knockdown (sh Kitl ) or overexpression ( Kitl OE). PLKO vector (shCtrl) and fluorescence overexpression ( Egfp OE) serves as the respective control. D, Representative images of CODEX staining (left) and quantification (right) for GFP (green) and CD31 (red) in normal pancreas from Fabp4-Cre or Kitl fl/fl ;Fabp4-Cre mouse model ( n = 3–4 mice per arm). Scale bar, 100 μm. E, Representative images of CODEX composite staining (left) and quantification (right) for CD45 (white) in normal pancreas from Fabp4-Cre or Kitl fl/fl ;Fabp4-Cre mouse model ( n = 4 mice per arm). Scale bar, 50 μm. F, Representative images of IHC staining for GFP (green), c-KIT receptor (KIT, red), and VIM (magenta) in healthy murine pancreas from Rosa26 mTmG/+ ;Fabp4-Cre mice. Scale bar, 10 μm. G, Representative images of IHC staining for CD31 (green) and c-KIT receptor (KIT, red) in healthy murine pancreas. Scale bar, 10 μm. H, Representative images of IHC staining for panCK (white) and c-KIT receptor (KIT, red) in healthy murine pancreas. Scale bar, 10 μm. I, Representative hematoxylin and eosin images between caerulein-treated Fabp4-Cre and Kitl fl/fl ;Fabp4-Cre mice. Scale bar, 100 μm. J, Representative images of IHC staining (left) for panCK (green) and amylase (red) between caerulein-treated Fabp4-Cre and Kitl fl/fl ;Fabp4-Cre mice. Scale bar, 10 μm. PanCK quantification on right ( n = 3 mice per arm). K, Representative IHC images (left) and quantification (right) for CD45 staining between caerulein-treated Fabp4-Cre and Kitl fl/fl ;Fabp4-Cre mice. Scale bar, 100 μm. L, Representative images (left) and quantification (right) for CD68 staining in caerulein-treated Fabp4-Cre and Kitl fl/fl ;Fabp4-Cre mouse pancreas. The same co-stained tissue section is shown in K and L . Scale bar, 100 μm. For comparisons of two groups, an unpaired t test was used. For comparisons of more than two groups with one independent variable, the one-way ANOVA test was used. Data are represented as mean ± SEM; *, P ≤ 0.05; **, P ≤ 0.01; ***, P ≤ 0.001; ****, P ≤ 0.0001; FC, fold changes; ns, not significant; panc area, pancreas.

    Techniques Used: Over Expression, RNA Sequencing, Quantitative RT-PCR, Expressing, Knockdown, Plasmid Preparation, Fluorescence, Control, Staining, Immunohistochemistry

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    Staining:

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    Article Snippet: CODEX staining and imaging was performed as described in user manual ( https://www.akoyabio.com/wp-content/uploads/2021/01/CODEX-User-Manual.pdf ). .. In brief, 5 μm FFPE pancreas sections were mounted onto 22 mm x 22 mm glass coverslips (Electron Microscopy Sciences) coated in 0.1% poly-L-lysine (Sigma) and stained with using CODEX Staining Kit (Akoya Biosciences). ..

    Article Title: Stromal KITL/SCF Maintains Pancreas Tissue Homeostasis and Restrains Tumor Progression
    Article Snippet: CODEX staining and imaging was performed as described in the user manual ( https://www.akoyabio.com/wp-content/uploads/2021/01/CODEX-User-Manual.pdf ). .. CODEX staining and imaging was performed as described in the user manual ( https://www.akoyabio.com/wp-content/uploads/2021/01/CODEX-User-Manual.pdf ). .. In brief, 5-μm FFPE pancreas sections were mounted onto 22 × 22 mm glass coverslips (Electron Microscopy Sciences) coated in 0.1% poly-L-lysine (Sigma-Aldrich) and stained with using CODEX Staining Kit (Akoya Biosciences).

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    Article Snippet: .. Multiplex-CODEX staining and imaging of the lungs of mice Barcode-conjugated antibody staining of tissue sections mounted on polylysine coated coverslips was performed using a commercially available CODEX Staining Kit for FFPE tissue using a commercial Akoya CODEX instrument (Akoya Biosciences, Marlborough, MA) with a Keyence BZ-X800 microscope plus Nikon PlanApo NA 0.75 objective. .. Images were acquired at 20X magnification and filter cubes for TRITC (550), CY5 (647), and CY7 (750) complementary fluorescent reporters and DAPI (Akoya Bio-sciences).

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    Formalin-fixed Paraffin-Embedded:

    Article Title: Gasdermin C promotes Stemness and Immune Evasion in Pancreatic Cancer via Pyroptosis-Independent Mechanism.
    Article Snippet: In addition, commercially available conjugated antibodies such as panCK (#232180, Akoya Biosciences) and Ki67 (#232179, Akoya Biosciences) were also utilized in the assays. .. For the staining of CODEX FFPE tissue, the CODEX Staining Kit (#232193, Akoya Biosciences) was used to accurately integrate fluorophores (reporters) into the oligonucleotide-antibody conjugates. .. This process included baking the coverslip at 65 °C for 30 minutes, followed by deparaffinization, rehydration, and epitope retrieval under high pressure using a Tris-EDTA Antigen Retrieval buffer solution with a pH of 9 (#abs9342, Absin).

    Article Title: Stromal KITL/SCF promotes pancreas tissue homeostasis and restrains tumor progression
    Article Snippet: CODEX staining and imaging was performed as described in user manual ( https://www.akoyabio.com/wp-content/uploads/2021/01/CODEX-User-Manual.pdf ). .. In brief, 5 μm FFPE pancreas sections were mounted onto 22 mm x 22 mm glass coverslips (Electron Microscopy Sciences) coated in 0.1% poly-L-lysine (Sigma) and stained with using CODEX Staining Kit (Akoya Biosciences). ..

    Article Title: Metabolic shifts in lipid utilization and reciprocal interactions within the lung metastatic niche of triple-negative breast cancer revealed by spatial multi-omics.
    Article Snippet: .. Multiplex-CODEX staining and imaging of the lungs of mice Barcode-conjugated antibody staining of tissue sections mounted on polylysine coated coverslips was performed using a commercially available CODEX Staining Kit for FFPE tissue using a commercial Akoya CODEX instrument (Akoya Biosciences, Marlborough, MA) with a Keyence BZ-X800 microscope plus Nikon PlanApo NA 0.75 objective. .. Images were acquired at 20X magnification and filter cubes for TRITC (550), CY5 (647), and CY7 (750) complementary fluorescent reporters and DAPI (Akoya Bio-sciences).

    Article Title: Metabolic shifts in lipid utilization and reciprocal interactions within the lung metastatic niche of triple-negative breast cancer revealed by spatial multi-omics
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    Article Title: Gasdermin C promotes Stemness and Immune Evasion in Pancreatic Cancer via Pyroptosis‐Independent Mechanism
    Article Snippet: In addition, commercially available conjugated antibodies such as panCK (#232180, Akoya Biosciences) and Ki67 (#232179, Akoya Biosciences) were also utilized in the assays. .. For the staining of CODEX FFPE tissue, the CODEX Staining Kit (#232193, Akoya Biosciences) was used to accurately integrate fluorophores (reporters) into the oligonucleotide‐antibody conjugates. .. This process included baking the coverslip at 65 °C for 30 minutes, followed by deparaffinization, rehydration, and epitope retrieval under high pressure using a Tris‐EDTA Antigen Retrieval buffer solution with a pH of 9 (#abs9342, Absin).

    Electron Microscopy:

    Article Title: Stromal KITL/SCF promotes pancreas tissue homeostasis and restrains tumor progression
    Article Snippet: CODEX staining and imaging was performed as described in user manual ( https://www.akoyabio.com/wp-content/uploads/2021/01/CODEX-User-Manual.pdf ). .. In brief, 5 μm FFPE pancreas sections were mounted onto 22 mm x 22 mm glass coverslips (Electron Microscopy Sciences) coated in 0.1% poly-L-lysine (Sigma) and stained with using CODEX Staining Kit (Akoya Biosciences). ..

    Imaging:

    Article Title: Stromal KITL/SCF Maintains Pancreas Tissue Homeostasis and Restrains Tumor Progression
    Article Snippet: CODEX staining and imaging was performed as described in the user manual ( https://www.akoyabio.com/wp-content/uploads/2021/01/CODEX-User-Manual.pdf ). .. CODEX staining and imaging was performed as described in the user manual ( https://www.akoyabio.com/wp-content/uploads/2021/01/CODEX-User-Manual.pdf ). .. In brief, 5-μm FFPE pancreas sections were mounted onto 22 × 22 mm glass coverslips (Electron Microscopy Sciences) coated in 0.1% poly-L-lysine (Sigma-Aldrich) and stained with using CODEX Staining Kit (Akoya Biosciences).

    Article Title: Metabolic shifts in lipid utilization and reciprocal interactions within the lung metastatic niche of triple-negative breast cancer revealed by spatial multi-omics.
    Article Snippet: .. Multiplex-CODEX staining and imaging of the lungs of mice Barcode-conjugated antibody staining of tissue sections mounted on polylysine coated coverslips was performed using a commercially available CODEX Staining Kit for FFPE tissue using a commercial Akoya CODEX instrument (Akoya Biosciences, Marlborough, MA) with a Keyence BZ-X800 microscope plus Nikon PlanApo NA 0.75 objective. .. Images were acquired at 20X magnification and filter cubes for TRITC (550), CY5 (647), and CY7 (750) complementary fluorescent reporters and DAPI (Akoya Bio-sciences).

    Multiplex Assay:

    Article Title: Metabolic shifts in lipid utilization and reciprocal interactions within the lung metastatic niche of triple-negative breast cancer revealed by spatial multi-omics.
    Article Snippet: .. Multiplex-CODEX staining and imaging of the lungs of mice Barcode-conjugated antibody staining of tissue sections mounted on polylysine coated coverslips was performed using a commercially available CODEX Staining Kit for FFPE tissue using a commercial Akoya CODEX instrument (Akoya Biosciences, Marlborough, MA) with a Keyence BZ-X800 microscope plus Nikon PlanApo NA 0.75 objective. .. Images were acquired at 20X magnification and filter cubes for TRITC (550), CY5 (647), and CY7 (750) complementary fluorescent reporters and DAPI (Akoya Bio-sciences).

    Microscopy:

    Article Title: Metabolic shifts in lipid utilization and reciprocal interactions within the lung metastatic niche of triple-negative breast cancer revealed by spatial multi-omics.
    Article Snippet: .. Multiplex-CODEX staining and imaging of the lungs of mice Barcode-conjugated antibody staining of tissue sections mounted on polylysine coated coverslips was performed using a commercially available CODEX Staining Kit for FFPE tissue using a commercial Akoya CODEX instrument (Akoya Biosciences, Marlborough, MA) with a Keyence BZ-X800 microscope plus Nikon PlanApo NA 0.75 objective. .. Images were acquired at 20X magnification and filter cubes for TRITC (550), CY5 (647), and CY7 (750) complementary fluorescent reporters and DAPI (Akoya Bio-sciences).

    Article Title: Metabolic shifts in lipid utilization and reciprocal interactions within the lung metastatic niche of triple-negative breast cancer revealed by spatial multi-omics
    Article Snippet: .. Barcode-conjugated antibody staining of tissue sections mounted on poly-lysine coated coverslips was performed using a commercially available CODEX Staining Kit for FFPE tissue using a commercial Akoya CODEX instrument (Akoya Biosciences, Marlborough, MA) with a Keyence BZ-X800 microscope plus Nikon PlanApo NA 0.75 objective. .. Images were acquired at 20X magnification and filter cubes for TRITC (550), CY5 (647), and CY7 (750) complementary fluorescent reporters and DAPI (Akoya Bio-sciences).



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    Technical Manufacturing Company codex staining kit
    KITL regulates PSC state and pancreas tissue homeostasis. A, Volcano plot of all upregulated, nonsignificant, and downregulated differentially expressed genes (DEG) as defined by the Wald test ( P .adj < 0.05 and log 2 FC > 1) from Kitl overexpression ( Kitl OE) ImPSC-1 bulk RNA-seq dataset with representative gene labels included. Data represent three biological repeats. B, Gene ontology (GO) analysis of upregulated and downregulated genes in immortalized PSCs (ImPSC-1) overexpressing Kitl . Top 10 enrichment categories ranked by adjusted P values plotted in each direction. C, qRT-PCR analysis of indicated DEGs in ImPSC-1 expressing stable Kitl knockdown (sh Kitl ) or overexpression ( Kitl OE). PLKO vector (shCtrl) and fluorescence overexpression ( Egfp OE) serves as the respective control. D, Representative images of <t>CODEX</t> <t>staining</t> (left) and quantification (right) for GFP (green) and CD31 (red) in normal pancreas from Fabp4-Cre or Kitl fl/fl ;Fabp4-Cre mouse model ( n = 3–4 mice per arm). Scale bar, 100 μm. E, Representative images of CODEX composite staining (left) and quantification (right) for CD45 (white) in normal pancreas from Fabp4-Cre or Kitl fl/fl ;Fabp4-Cre mouse model ( n = 4 mice per arm). Scale bar, 50 μm. F, Representative images of IHC staining for GFP (green), c-KIT receptor (KIT, red), and VIM (magenta) in healthy murine pancreas from Rosa26 mTmG/+ ;Fabp4-Cre mice. Scale bar, 10 μm. G, Representative images of IHC staining for CD31 (green) and c-KIT receptor (KIT, red) in healthy murine pancreas. Scale bar, 10 μm. H, Representative images of IHC staining for panCK (white) and c-KIT receptor (KIT, red) in healthy murine pancreas. Scale bar, 10 μm. I, Representative hematoxylin and eosin images between caerulein-treated Fabp4-Cre and Kitl fl/fl ;Fabp4-Cre mice. Scale bar, 100 μm. J, Representative images of IHC staining (left) for panCK (green) and amylase (red) between caerulein-treated Fabp4-Cre and Kitl fl/fl ;Fabp4-Cre mice. Scale bar, 10 μm. PanCK quantification on right ( n = 3 mice per arm). K, Representative IHC images (left) and quantification (right) for CD45 staining between caerulein-treated Fabp4-Cre and Kitl fl/fl ;Fabp4-Cre mice. Scale bar, 100 μm. L, Representative images (left) and quantification (right) for CD68 staining in caerulein-treated Fabp4-Cre and Kitl fl/fl ;Fabp4-Cre mouse pancreas. The same co-stained tissue section is shown in K and L . Scale bar, 100 μm. For comparisons of two groups, an unpaired t test was used. For comparisons of more than two groups with one independent variable, the one-way ANOVA test was used. Data are represented as mean ± SEM; *, P ≤ 0.05; **, P ≤ 0.01; ***, P ≤ 0.001; ****, P ≤ 0.0001; FC, fold changes; ns, not significant; panc area, pancreas.
    Codex Staining Kit, supplied by Technical Manufacturing Company, 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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    KITL regulates PSC state and pancreas tissue homeostasis. A, Volcano plot of all upregulated, nonsignificant, and downregulated differentially expressed genes (DEG) as defined by the Wald test ( P .adj < 0.05 and log 2 FC > 1) from Kitl overexpression ( Kitl OE) ImPSC-1 bulk RNA-seq dataset with representative gene labels included. Data represent three biological repeats. B, Gene ontology (GO) analysis of upregulated and downregulated genes in immortalized PSCs (ImPSC-1) overexpressing Kitl . Top 10 enrichment categories ranked by adjusted P values plotted in each direction. C, qRT-PCR analysis of indicated DEGs in ImPSC-1 expressing stable Kitl knockdown (sh Kitl ) or overexpression ( Kitl OE). PLKO vector (shCtrl) and fluorescence overexpression ( Egfp OE) serves as the respective control. D, Representative images of CODEX staining (left) and quantification (right) for GFP (green) and CD31 (red) in normal pancreas from Fabp4-Cre or Kitl fl/fl ;Fabp4-Cre mouse model ( n = 3–4 mice per arm). Scale bar, 100 μm. E, Representative images of CODEX composite staining (left) and quantification (right) for CD45 (white) in normal pancreas from Fabp4-Cre or Kitl fl/fl ;Fabp4-Cre mouse model ( n = 4 mice per arm). Scale bar, 50 μm. F, Representative images of IHC staining for GFP (green), c-KIT receptor (KIT, red), and VIM (magenta) in healthy murine pancreas from Rosa26 mTmG/+ ;Fabp4-Cre mice. Scale bar, 10 μm. G, Representative images of IHC staining for CD31 (green) and c-KIT receptor (KIT, red) in healthy murine pancreas. Scale bar, 10 μm. H, Representative images of IHC staining for panCK (white) and c-KIT receptor (KIT, red) in healthy murine pancreas. Scale bar, 10 μm. I, Representative hematoxylin and eosin images between caerulein-treated Fabp4-Cre and Kitl fl/fl ;Fabp4-Cre mice. Scale bar, 100 μm. J, Representative images of IHC staining (left) for panCK (green) and amylase (red) between caerulein-treated Fabp4-Cre and Kitl fl/fl ;Fabp4-Cre mice. Scale bar, 10 μm. PanCK quantification on right ( n = 3 mice per arm). K, Representative IHC images (left) and quantification (right) for CD45 staining between caerulein-treated Fabp4-Cre and Kitl fl/fl ;Fabp4-Cre mice. Scale bar, 100 μm. L, Representative images (left) and quantification (right) for CD68 staining in caerulein-treated Fabp4-Cre and Kitl fl/fl ;Fabp4-Cre mouse pancreas. The same co-stained tissue section is shown in K and L . Scale bar, 100 μm. For comparisons of two groups, an unpaired t test was used. For comparisons of more than two groups with one independent variable, the one-way ANOVA test was used. Data are represented as mean ± SEM; *, P ≤ 0.05; **, P ≤ 0.01; ***, P ≤ 0.001; ****, P ≤ 0.0001; FC, fold changes; ns, not significant; panc area, pancreas.

    Journal: Cancer Discovery

    Article Title: Stromal KITL/SCF Maintains Pancreas Tissue Homeostasis and Restrains Tumor Progression

    doi: 10.1158/2159-8290.CD-24-1079

    Figure Lengend Snippet: KITL regulates PSC state and pancreas tissue homeostasis. A, Volcano plot of all upregulated, nonsignificant, and downregulated differentially expressed genes (DEG) as defined by the Wald test ( P .adj < 0.05 and log 2 FC > 1) from Kitl overexpression ( Kitl OE) ImPSC-1 bulk RNA-seq dataset with representative gene labels included. Data represent three biological repeats. B, Gene ontology (GO) analysis of upregulated and downregulated genes in immortalized PSCs (ImPSC-1) overexpressing Kitl . Top 10 enrichment categories ranked by adjusted P values plotted in each direction. C, qRT-PCR analysis of indicated DEGs in ImPSC-1 expressing stable Kitl knockdown (sh Kitl ) or overexpression ( Kitl OE). PLKO vector (shCtrl) and fluorescence overexpression ( Egfp OE) serves as the respective control. D, Representative images of CODEX staining (left) and quantification (right) for GFP (green) and CD31 (red) in normal pancreas from Fabp4-Cre or Kitl fl/fl ;Fabp4-Cre mouse model ( n = 3–4 mice per arm). Scale bar, 100 μm. E, Representative images of CODEX composite staining (left) and quantification (right) for CD45 (white) in normal pancreas from Fabp4-Cre or Kitl fl/fl ;Fabp4-Cre mouse model ( n = 4 mice per arm). Scale bar, 50 μm. F, Representative images of IHC staining for GFP (green), c-KIT receptor (KIT, red), and VIM (magenta) in healthy murine pancreas from Rosa26 mTmG/+ ;Fabp4-Cre mice. Scale bar, 10 μm. G, Representative images of IHC staining for CD31 (green) and c-KIT receptor (KIT, red) in healthy murine pancreas. Scale bar, 10 μm. H, Representative images of IHC staining for panCK (white) and c-KIT receptor (KIT, red) in healthy murine pancreas. Scale bar, 10 μm. I, Representative hematoxylin and eosin images between caerulein-treated Fabp4-Cre and Kitl fl/fl ;Fabp4-Cre mice. Scale bar, 100 μm. J, Representative images of IHC staining (left) for panCK (green) and amylase (red) between caerulein-treated Fabp4-Cre and Kitl fl/fl ;Fabp4-Cre mice. Scale bar, 10 μm. PanCK quantification on right ( n = 3 mice per arm). K, Representative IHC images (left) and quantification (right) for CD45 staining between caerulein-treated Fabp4-Cre and Kitl fl/fl ;Fabp4-Cre mice. Scale bar, 100 μm. L, Representative images (left) and quantification (right) for CD68 staining in caerulein-treated Fabp4-Cre and Kitl fl/fl ;Fabp4-Cre mouse pancreas. The same co-stained tissue section is shown in K and L . Scale bar, 100 μm. For comparisons of two groups, an unpaired t test was used. For comparisons of more than two groups with one independent variable, the one-way ANOVA test was used. Data are represented as mean ± SEM; *, P ≤ 0.05; **, P ≤ 0.01; ***, P ≤ 0.001; ****, P ≤ 0.0001; FC, fold changes; ns, not significant; panc area, pancreas.

    Article Snippet: In brief, 5-μm FFPE pancreas sections were mounted onto 22 × 22 mm glass coverslips (Electron Microscopy Sciences) coated in 0.1% poly-L-lysine (Sigma-Aldrich) and stained with using CODEX Staining Kit (Akoya Biosciences).

    Techniques: Over Expression, RNA Sequencing, Quantitative RT-PCR, Expressing, Knockdown, Plasmid Preparation, Fluorescence, Control, Staining, Immunohistochemistry