Journal: bioRxiv

Article Title: Restoration of Keratinocyte Homeostasis Drives Resolution of Skin Inflammation

doi: 10.64898/2026.03.04.708224

Figure Lengend Snippet: a, Experimental schematic for topical siRNA-mediated gene silencing in KC-Tie2 mice. Non-targeting control siRNA or gene-specific siRNAs targeting Cacna2d1 , Serpinb3b , or Slc25a5 were applied topically to ear skin at the indicated time points, with tissue collection on day 14. b, Representative hematoxylin and eosin–stained sections of ear skin following treatment with control siRNA or gene-specific siRNAs. c, Heatmap summarizing changes in epidermal acanthosis and expression of IL-23/IL-17-axis-associated genes ( Il23a , Ccl20 , S100a8 , Il17a , Tnf , Il17c ) following gene-specific knockdown. Values are shown as log₂ fold change relative to control siRNA-treated skin. Acanthosis is coded as increased or decreased relative to control. d, Correlation heatmap relating each targeted gene to transcriptional programs identified by bulk RNA-seq pathway analyses, including fatty acid and lipid metabolism, epidermal barrier and junctional organization, IL-17-associated inflammatory signaling, epidermal differentiation and cornified envelope formation, cholesterol and sterol biosynthesis, and neutrophil activation/degranulation. Quantitative log₂ fold-change values underlying the cytokine and alarmin heatmap in c are provided in Supplementary Table 29.

Article Snippet: Mouse primers (ThermoFisher Scientific) used in this study were: Il23a , Mm01160011_g1; Il17a , Mm00439618_m1; Il17c, Mm00521397_m1, Ccl20 , Mm00444228_m1; S100a8 , Mm00495696_g1; Tnf , Mm00443260_g1, Cacna2d1 , Mm00486607_m1; Serpinb3b , Mm03032256_uH; Slc25a5 , Mm00846873_g1; and Gapdh , Mm99999915_g1.

Techniques: Control, Staining, Expressing, Knockdown, RNA Sequencing, Activation Assay

Journal: bioRxiv

Article Title: Mutant KRAS promotes NF-κB driven CCL20 chemokine expression in pancreatic ductal adenocarcinoma

doi: 10.64898/2026.04.13.717530

Figure Lengend Snippet: A Bulk sequencing data from TCGA (left) and GTEx (right) showing CCL20 expression. Gastrointestinal tissues prominently express CCL20 while other epithelial tissues expressed little if any CCL20. B Expression of CCL20 by the different cell types found in primary tumors from a PDAC scRNA atlas . C CCL20 expression from a scRNA dataset healthy human colon (H) and colorectal cancer . D Expression of CCL20 by different cells found in a single nucleus RNA sequencing dataset of chronic pancreatitis biopsies . E CCR6, CCL20, IL-17A, and IL-23A in a scRNA datasets of CD45+ cells from human chronic pancreatitis or healthy patients . Pancreatitis was classified as hereditary if there were mutations present in PRSS1 gene. F Expression of CCL20 in a scRNA dataset of human ulcerative colitis (UC) and Crohn’s disease (CD) . G Differentially expressed chemokines found using MAST from a scRNA atlas of PDAC .

Article Snippet: The ELISAs were performed using the human CCL20 ELISA (R&D Systems, Minneapolis, MN, #DY360) or mouse CCL20 ELISA (R&D Systems #DY760) following the manufacturer’s protocol.

Techniques: Sequencing, Expressing, RNA Sequencing

Journal: bioRxiv

Article Title: Mutant KRAS promotes NF-κB driven CCL20 chemokine expression in pancreatic ductal adenocarcinoma

doi: 10.64898/2026.04.13.717530

Figure Lengend Snippet: A Representative PCR analyses of CCL20 and CCR6 transcript expression across multiple human and B mouse PDAC cell lines. Data representative of 3 individual analyses. C Quantification of the CCL20 staining images. 3-7 fields of view were quantified per patient, with five patients per disease state . D Representative immunostaining for CCL20 (red) or DAPI-stained cell nuclei (blue) (top) with parallel sections stained with H&E (bottom) in both healthy human pancreas or E human PDAC samples. * denotes p ≤ 0.05, Student’s unpaired t-test. Size bar = 100 μm.

Article Snippet: The ELISAs were performed using the human CCL20 ELISA (R&D Systems, Minneapolis, MN, #DY360) or mouse CCL20 ELISA (R&D Systems #DY760) following the manufacturer’s protocol.

Techniques: Expressing, Staining, Immunostaining

Journal: bioRxiv

Article Title: Mutant KRAS promotes NF-κB driven CCL20 chemokine expression in pancreatic ductal adenocarcinoma

doi: 10.64898/2026.04.13.717530

Figure Lengend Snippet: A Gene sets listed in Supplementary Data 7 were correlated with CCL20 expression in ductal cells from a scRNA dataset of human PDAC and B colorectal cancer . Values shown are Pearson’s correlation coefficients, all p values = 0. C TCGA PDAC patients and D colorectal cancer patients sorted by SBS1 (replicative age) signatures, with comparisons of their normalized CCL20 expression. E TCGA PDAC patients sorted by their CN1 (diploid) and F CN2 (tetraploid) signatures using data from previously reported . Similar analysis for colorectal cancer patients can be seen in G and H . I Normalized CCL20 methylation values (probe cg21643045) from all GI cancers (cancer types in the top row of ) sorted by CCL20 expression. J CCL20 high patients (relative to patients of same cancer type) with their CCL20 normalized methylation values plotted. Non-GI mucosal cancers are bladder, cervical, head and neck, both lung types, and both uterine tumor types. Panel C-I Student’s unpaired t -test was used with p values plotted. Panel J one-way ANOVA with Tukey’s test for multiple comparisons, * denotes p ≤ 0.05.

Article Snippet: The ELISAs were performed using the human CCL20 ELISA (R&D Systems, Minneapolis, MN, #DY360) or mouse CCL20 ELISA (R&D Systems #DY760) following the manufacturer’s protocol.

Techniques: Expressing, Methylation

Journal: bioRxiv

Article Title: Mutant KRAS promotes NF-κB driven CCL20 chemokine expression in pancreatic ductal adenocarcinoma

doi: 10.64898/2026.04.13.717530

Figure Lengend Snippet: A The top three transcription factors inferred using CollecTRI from all pancreatic exocrine ductal cells in a single cell RNA dataset sorted by CCL20 expression. B Inferred transcription factors from tumor cells from a CRC dataset sorted by CCL20 expression. C qPCR and D ELISA for CCL20 transcript expression and protein secretion by human PANC-1 cell line. E qPCR for CXCL2 by the PANC-1 cell line. ND = not detected by qPCR. ns, not significant. F qPCR for CCL20 and G CXCL2 expression by the patient-derived 339 cell line possessing a KRAS G12V mutation. H qPCR and I ELISA for CCL20 in the murine KPC FC1242 stably expressing Firefly luciferase PDAC cell line. Treatments were for 24 hours. Concentrations of the individual treatments were: Erlotinib 500 nM, regorafenib 500 nM, MRTX1133 500 nM, gemcitabine 100 nM, TNF 20 ng/mL with 10 ng/mL of IL-1, and RMC-7977 300 nM. For resistant cell lines (denoted by -R), resistance was considered established when cells survived in full growth medium supplemented with 10 μM of the denoted chemotherapy drug. One-way ANOVA was used for statistical analysis with Dunnett’s test for multiple comparisons used to compare to the untreated cell line.

Article Snippet: The ELISAs were performed using the human CCL20 ELISA (R&D Systems, Minneapolis, MN, #DY360) or mouse CCL20 ELISA (R&D Systems #DY760) following the manufacturer’s protocol.

Techniques: Single Cell, Expressing, Enzyme-linked Immunosorbent Assay, Derivative Assay, Mutagenesis, Stable Transfection, Luciferase

Journal: bioRxiv

Article Title: Mutant KRAS promotes NF-κB driven CCL20 chemokine expression in pancreatic ductal adenocarcinoma

doi: 10.64898/2026.04.13.717530

Figure Lengend Snippet: A Mean fluorescence intensity (MFI) of anti-phospho-p65-PE immunostained FC1245 and RMC-7977 (2 μM)-resistant FC1245 PDAC cells. * denotes p ≤ 0.05, Student’s unpaired t -test. B Expression of CCL20 in a single cell RNA dataset of pancreatic ductal cells obtained from mice with a Ptf1a specific tamoxifen inducible Kras -G12D. The time listed on the y-axis is time from when the initial tamoxifen was given. C The number of wild-type C57BL6 splenocytes migrated in a chemotaxis assay using conditioned media from the listed FC1242 PDAC cell line. The conditioned media was obtained from cells outlined in . One-way ANOVA was used for statistical analysis with Dunnett’s test for multiple comparisons used to compare to the untreated cell line D Treatment schema for mice treated implanted with orthotopic KPC1199 tumors and treated with either vehicle PBS or 100 μg CCL20LD. E The mass of the tumors from the experiment outlined in . F CD11c+ MHCII+ dendritic cells, G CD11b+ F4/80+ macrophages, and H MHCII high macrophages (as a percentage of macrophages) in the harvested tumors. * denotes p ≤ 0.05, Student’s unpaired t -test.

Article Snippet: The ELISAs were performed using the human CCL20 ELISA (R&D Systems, Minneapolis, MN, #DY360) or mouse CCL20 ELISA (R&D Systems #DY760) following the manufacturer’s protocol.

Techniques: Fluorescence, Expressing, Single Cell, Chemotaxis Assay

Journal: bioRxiv

Article Title: Mutant KRAS promotes NF-κB driven CCL20 chemokine expression in pancreatic ductal adenocarcinoma

doi: 10.64898/2026.04.13.717530

Figure Lengend Snippet: A Graphical abstract summarizing the findings. CCL20 is not expressed in the healthy pancreas. In pancreatitis, it is expressed the lymphocytic infiltrate. In PanINs, CCL20 begins to be expressed by the ductal epithelium. In PDAC, CCL20 expression is further increased by the ductal tumor cells and infiltrating myeloid cells. CCL20 expression is lost with resistance to KRAS inhibitors, along with decreased phosphorylation of RELA , and is limited to the basal subtype. Inhibition of CCL20-CCR6 signaling increased levels of antigen presenting cells, including dendritic cells and MHCII-high macrophages within the tumor parenchyma.

Article Snippet: The ELISAs were performed using the human CCL20 ELISA (R&D Systems, Minneapolis, MN, #DY360) or mouse CCL20 ELISA (R&D Systems #DY760) following the manufacturer’s protocol.

Techniques: Expressing, Phospho-proteomics, Inhibition

Journal: bioRxiv

Article Title: Mutant KRAS promotes NF-κB driven CCL20 chemokine expression in pancreatic ductal adenocarcinoma

doi: 10.64898/2026.04.13.717530

Figure Lengend Snippet: A Bulk sequencing data from TCGA (left) and GTEx (right) showing CCL20 expression. Gastrointestinal tissues prominently express CCL20 while other epithelial tissues expressed little if any CCL20. B Expression of CCL20 by the different cell types found in primary tumors from a PDAC scRNA atlas . C CCL20 expression from a scRNA dataset healthy human colon (H) and colorectal cancer . D Expression of CCL20 by different cells found in a single nucleus RNA sequencing dataset of chronic pancreatitis biopsies . E CCR6, CCL20, IL-17A, and IL-23A in a scRNA datasets of CD45+ cells from human chronic pancreatitis or healthy patients . Pancreatitis was classified as hereditary if there were mutations present in PRSS1 gene. F Expression of CCL20 in a scRNA dataset of human ulcerative colitis (UC) and Crohn’s disease (CD) . G Differentially expressed chemokines found using MAST from a scRNA atlas of PDAC .

Article Snippet: The ELISAs were performed using the human CCL20 ELISA (R&D Systems, Minneapolis, MN, #DY360) or mouse CCL20 ELISA (R&D Systems #DY760) following the manufacturer’s protocol.

Techniques: Sequencing, Expressing, RNA Sequencing

Journal: bioRxiv

Article Title: Mutant KRAS promotes NF-κB driven CCL20 chemokine expression in pancreatic ductal adenocarcinoma

doi: 10.64898/2026.04.13.717530

Figure Lengend Snippet: A Representative PCR analyses of CCL20 and CCR6 transcript expression across multiple human and B mouse PDAC cell lines. Data representative of 3 individual analyses. C Quantification of the CCL20 staining images. 3-7 fields of view were quantified per patient, with five patients per disease state . D Representative immunostaining for CCL20 (red) or DAPI-stained cell nuclei (blue) (top) with parallel sections stained with H&E (bottom) in both healthy human pancreas or E human PDAC samples. * denotes p ≤ 0.05, Student’s unpaired t-test. Size bar = 100 μm.

Article Snippet: The ELISAs were performed using the human CCL20 ELISA (R&D Systems, Minneapolis, MN, #DY360) or mouse CCL20 ELISA (R&D Systems #DY760) following the manufacturer’s protocol.

Techniques: Expressing, Staining, Immunostaining

Journal: bioRxiv

Article Title: Mutant KRAS promotes NF-κB driven CCL20 chemokine expression in pancreatic ductal adenocarcinoma

doi: 10.64898/2026.04.13.717530

Figure Lengend Snippet: A Gene sets listed in Supplementary Data 7 were correlated with CCL20 expression in ductal cells from a scRNA dataset of human PDAC and B colorectal cancer . Values shown are Pearson’s correlation coefficients, all p values = 0. C TCGA PDAC patients and D colorectal cancer patients sorted by SBS1 (replicative age) signatures, with comparisons of their normalized CCL20 expression. E TCGA PDAC patients sorted by their CN1 (diploid) and F CN2 (tetraploid) signatures using data from previously reported . Similar analysis for colorectal cancer patients can be seen in G and H . I Normalized CCL20 methylation values (probe cg21643045) from all GI cancers (cancer types in the top row of ) sorted by CCL20 expression. J CCL20 high patients (relative to patients of same cancer type) with their CCL20 normalized methylation values plotted. Non-GI mucosal cancers are bladder, cervical, head and neck, both lung types, and both uterine tumor types. Panel C-I Student’s unpaired t -test was used with p values plotted. Panel J one-way ANOVA with Tukey’s test for multiple comparisons, * denotes p ≤ 0.05.

Article Snippet: The ELISAs were performed using the human CCL20 ELISA (R&D Systems, Minneapolis, MN, #DY360) or mouse CCL20 ELISA (R&D Systems #DY760) following the manufacturer’s protocol.

Techniques: Expressing, Methylation

Journal: bioRxiv

Article Title: Mutant KRAS promotes NF-κB driven CCL20 chemokine expression in pancreatic ductal adenocarcinoma

doi: 10.64898/2026.04.13.717530

Figure Lengend Snippet: A The top three transcription factors inferred using CollecTRI from all pancreatic exocrine ductal cells in a single cell RNA dataset sorted by CCL20 expression. B Inferred transcription factors from tumor cells from a CRC dataset sorted by CCL20 expression. C qPCR and D ELISA for CCL20 transcript expression and protein secretion by human PANC-1 cell line. E qPCR for CXCL2 by the PANC-1 cell line. ND = not detected by qPCR. ns, not significant. F qPCR for CCL20 and G CXCL2 expression by the patient-derived 339 cell line possessing a KRAS G12V mutation. H qPCR and I ELISA for CCL20 in the murine KPC FC1242 stably expressing Firefly luciferase PDAC cell line. Treatments were for 24 hours. Concentrations of the individual treatments were: Erlotinib 500 nM, regorafenib 500 nM, MRTX1133 500 nM, gemcitabine 100 nM, TNF 20 ng/mL with 10 ng/mL of IL-1, and RMC-7977 300 nM. For resistant cell lines (denoted by -R), resistance was considered established when cells survived in full growth medium supplemented with 10 μM of the denoted chemotherapy drug. One-way ANOVA was used for statistical analysis with Dunnett’s test for multiple comparisons used to compare to the untreated cell line.

Article Snippet: The ELISAs were performed using the human CCL20 ELISA (R&D Systems, Minneapolis, MN, #DY360) or mouse CCL20 ELISA (R&D Systems #DY760) following the manufacturer’s protocol.

Techniques: Single Cell, Expressing, Enzyme-linked Immunosorbent Assay, Derivative Assay, Mutagenesis, Stable Transfection, Luciferase

Journal: bioRxiv

Article Title: Mutant KRAS promotes NF-κB driven CCL20 chemokine expression in pancreatic ductal adenocarcinoma

doi: 10.64898/2026.04.13.717530

Figure Lengend Snippet: A Mean fluorescence intensity (MFI) of anti-phospho-p65-PE immunostained FC1245 and RMC-7977 (2 μM)-resistant FC1245 PDAC cells. * denotes p ≤ 0.05, Student’s unpaired t -test. B Expression of CCL20 in a single cell RNA dataset of pancreatic ductal cells obtained from mice with a Ptf1a specific tamoxifen inducible Kras -G12D. The time listed on the y-axis is time from when the initial tamoxifen was given. C The number of wild-type C57BL6 splenocytes migrated in a chemotaxis assay using conditioned media from the listed FC1242 PDAC cell line. The conditioned media was obtained from cells outlined in . One-way ANOVA was used for statistical analysis with Dunnett’s test for multiple comparisons used to compare to the untreated cell line D Treatment schema for mice treated implanted with orthotopic KPC1199 tumors and treated with either vehicle PBS or 100 μg CCL20LD. E The mass of the tumors from the experiment outlined in . F CD11c+ MHCII+ dendritic cells, G CD11b+ F4/80+ macrophages, and H MHCII high macrophages (as a percentage of macrophages) in the harvested tumors. * denotes p ≤ 0.05, Student’s unpaired t -test.

Article Snippet: The ELISAs were performed using the human CCL20 ELISA (R&D Systems, Minneapolis, MN, #DY360) or mouse CCL20 ELISA (R&D Systems #DY760) following the manufacturer’s protocol.

Techniques: Fluorescence, Expressing, Single Cell, Chemotaxis Assay

Journal: bioRxiv

Article Title: Mutant KRAS promotes NF-κB driven CCL20 chemokine expression in pancreatic ductal adenocarcinoma

doi: 10.64898/2026.04.13.717530

Figure Lengend Snippet: A Graphical abstract summarizing the findings. CCL20 is not expressed in the healthy pancreas. In pancreatitis, it is expressed the lymphocytic infiltrate. In PanINs, CCL20 begins to be expressed by the ductal epithelium. In PDAC, CCL20 expression is further increased by the ductal tumor cells and infiltrating myeloid cells. CCL20 expression is lost with resistance to KRAS inhibitors, along with decreased phosphorylation of RELA , and is limited to the basal subtype. Inhibition of CCL20-CCR6 signaling increased levels of antigen presenting cells, including dendritic cells and MHCII-high macrophages within the tumor parenchyma.

Article Snippet: The ELISAs were performed using the human CCL20 ELISA (R&D Systems, Minneapolis, MN, #DY360) or mouse CCL20 ELISA (R&D Systems #DY760) following the manufacturer’s protocol.

Techniques: Expressing, Phospho-proteomics, Inhibition

Journal: bioRxiv

Article Title: Mutant KRAS promotes NF-κB driven CCL20 chemokine expression in pancreatic ductal adenocarcinoma

doi: 10.64898/2026.04.13.717530

Figure Lengend Snippet: A Bulk sequencing data from TCGA (left) and GTEx (right) showing CCL20 expression. Gastrointestinal tissues prominently express CCL20 while other epithelial tissues expressed little if any CCL20. B Expression of CCL20 by the different cell types found in primary tumors from a PDAC scRNA atlas . C CCL20 expression from a scRNA dataset healthy human colon (H) and colorectal cancer . D Expression of CCL20 by different cells found in a single nucleus RNA sequencing dataset of chronic pancreatitis biopsies . E CCR6, CCL20, IL-17A, and IL-23A in a scRNA datasets of CD45+ cells from human chronic pancreatitis or healthy patients . Pancreatitis was classified as hereditary if there were mutations present in PRSS1 gene. F Expression of CCL20 in a scRNA dataset of human ulcerative colitis (UC) and Crohn’s disease (CD) . G Differentially expressed chemokines found using MAST from a scRNA atlas of PDAC .

Article Snippet: Samples were deparaffinized, blocked with 2% (v/v) goat serum (ThermoFisher #31872), stained with mouse anti-human CCL20 (R&D systems MAB360) at a concentration of 1:100 and goat anti-mouse IgG Alexa fluor 647 (ThermoFisher #A48289) at a concentration of 1:500.

Techniques: Sequencing, Expressing, RNA Sequencing

Journal: bioRxiv

Article Title: Mutant KRAS promotes NF-κB driven CCL20 chemokine expression in pancreatic ductal adenocarcinoma

doi: 10.64898/2026.04.13.717530

Figure Lengend Snippet: A Representative PCR analyses of CCL20 and CCR6 transcript expression across multiple human and B mouse PDAC cell lines. Data representative of 3 individual analyses. C Quantification of the CCL20 staining images. 3-7 fields of view were quantified per patient, with five patients per disease state . D Representative immunostaining for CCL20 (red) or DAPI-stained cell nuclei (blue) (top) with parallel sections stained with H&E (bottom) in both healthy human pancreas or E human PDAC samples. * denotes p ≤ 0.05, Student’s unpaired t-test. Size bar = 100 μm.

Article Snippet: Samples were deparaffinized, blocked with 2% (v/v) goat serum (ThermoFisher #31872), stained with mouse anti-human CCL20 (R&D systems MAB360) at a concentration of 1:100 and goat anti-mouse IgG Alexa fluor 647 (ThermoFisher #A48289) at a concentration of 1:500.

Techniques: Expressing, Staining, Immunostaining

Journal: bioRxiv

Article Title: Mutant KRAS promotes NF-κB driven CCL20 chemokine expression in pancreatic ductal adenocarcinoma

doi: 10.64898/2026.04.13.717530

Figure Lengend Snippet: A Gene sets listed in Supplementary Data 7 were correlated with CCL20 expression in ductal cells from a scRNA dataset of human PDAC and B colorectal cancer . Values shown are Pearson’s correlation coefficients, all p values = 0. C TCGA PDAC patients and D colorectal cancer patients sorted by SBS1 (replicative age) signatures, with comparisons of their normalized CCL20 expression. E TCGA PDAC patients sorted by their CN1 (diploid) and F CN2 (tetraploid) signatures using data from previously reported . Similar analysis for colorectal cancer patients can be seen in G and H . I Normalized CCL20 methylation values (probe cg21643045) from all GI cancers (cancer types in the top row of ) sorted by CCL20 expression. J CCL20 high patients (relative to patients of same cancer type) with their CCL20 normalized methylation values plotted. Non-GI mucosal cancers are bladder, cervical, head and neck, both lung types, and both uterine tumor types. Panel C-I Student’s unpaired t -test was used with p values plotted. Panel J one-way ANOVA with Tukey’s test for multiple comparisons, * denotes p ≤ 0.05.

Article Snippet: Samples were deparaffinized, blocked with 2% (v/v) goat serum (ThermoFisher #31872), stained with mouse anti-human CCL20 (R&D systems MAB360) at a concentration of 1:100 and goat anti-mouse IgG Alexa fluor 647 (ThermoFisher #A48289) at a concentration of 1:500.

Techniques: Expressing, Methylation

Journal: bioRxiv

Article Title: Mutant KRAS promotes NF-κB driven CCL20 chemokine expression in pancreatic ductal adenocarcinoma

doi: 10.64898/2026.04.13.717530

Figure Lengend Snippet: A The top three transcription factors inferred using CollecTRI from all pancreatic exocrine ductal cells in a single cell RNA dataset sorted by CCL20 expression. B Inferred transcription factors from tumor cells from a CRC dataset sorted by CCL20 expression. C qPCR and D ELISA for CCL20 transcript expression and protein secretion by human PANC-1 cell line. E qPCR for CXCL2 by the PANC-1 cell line. ND = not detected by qPCR. ns, not significant. F qPCR for CCL20 and G CXCL2 expression by the patient-derived 339 cell line possessing a KRAS G12V mutation. H qPCR and I ELISA for CCL20 in the murine KPC FC1242 stably expressing Firefly luciferase PDAC cell line. Treatments were for 24 hours. Concentrations of the individual treatments were: Erlotinib 500 nM, regorafenib 500 nM, MRTX1133 500 nM, gemcitabine 100 nM, TNF 20 ng/mL with 10 ng/mL of IL-1, and RMC-7977 300 nM. For resistant cell lines (denoted by -R), resistance was considered established when cells survived in full growth medium supplemented with 10 μM of the denoted chemotherapy drug. One-way ANOVA was used for statistical analysis with Dunnett’s test for multiple comparisons used to compare to the untreated cell line.

Article Snippet: Samples were deparaffinized, blocked with 2% (v/v) goat serum (ThermoFisher #31872), stained with mouse anti-human CCL20 (R&D systems MAB360) at a concentration of 1:100 and goat anti-mouse IgG Alexa fluor 647 (ThermoFisher #A48289) at a concentration of 1:500.

Techniques: Single Cell, Expressing, Enzyme-linked Immunosorbent Assay, Derivative Assay, Mutagenesis, Stable Transfection, Luciferase

Journal: bioRxiv

Article Title: Mutant KRAS promotes NF-κB driven CCL20 chemokine expression in pancreatic ductal adenocarcinoma

doi: 10.64898/2026.04.13.717530

Figure Lengend Snippet: A Mean fluorescence intensity (MFI) of anti-phospho-p65-PE immunostained FC1245 and RMC-7977 (2 μM)-resistant FC1245 PDAC cells. * denotes p ≤ 0.05, Student’s unpaired t -test. B Expression of CCL20 in a single cell RNA dataset of pancreatic ductal cells obtained from mice with a Ptf1a specific tamoxifen inducible Kras -G12D. The time listed on the y-axis is time from when the initial tamoxifen was given. C The number of wild-type C57BL6 splenocytes migrated in a chemotaxis assay using conditioned media from the listed FC1242 PDAC cell line. The conditioned media was obtained from cells outlined in . One-way ANOVA was used for statistical analysis with Dunnett’s test for multiple comparisons used to compare to the untreated cell line D Treatment schema for mice treated implanted with orthotopic KPC1199 tumors and treated with either vehicle PBS or 100 μg CCL20LD. E The mass of the tumors from the experiment outlined in . F CD11c+ MHCII+ dendritic cells, G CD11b+ F4/80+ macrophages, and H MHCII high macrophages (as a percentage of macrophages) in the harvested tumors. * denotes p ≤ 0.05, Student’s unpaired t -test.

Article Snippet: Samples were deparaffinized, blocked with 2% (v/v) goat serum (ThermoFisher #31872), stained with mouse anti-human CCL20 (R&D systems MAB360) at a concentration of 1:100 and goat anti-mouse IgG Alexa fluor 647 (ThermoFisher #A48289) at a concentration of 1:500.

Techniques: Fluorescence, Expressing, Single Cell, Chemotaxis Assay

Journal: bioRxiv

Article Title: Mutant KRAS promotes NF-κB driven CCL20 chemokine expression in pancreatic ductal adenocarcinoma

doi: 10.64898/2026.04.13.717530

Figure Lengend Snippet: A Graphical abstract summarizing the findings. CCL20 is not expressed in the healthy pancreas. In pancreatitis, it is expressed the lymphocytic infiltrate. In PanINs, CCL20 begins to be expressed by the ductal epithelium. In PDAC, CCL20 expression is further increased by the ductal tumor cells and infiltrating myeloid cells. CCL20 expression is lost with resistance to KRAS inhibitors, along with decreased phosphorylation of RELA , and is limited to the basal subtype. Inhibition of CCL20-CCR6 signaling increased levels of antigen presenting cells, including dendritic cells and MHCII-high macrophages within the tumor parenchyma.

Article Snippet: Samples were deparaffinized, blocked with 2% (v/v) goat serum (ThermoFisher #31872), stained with mouse anti-human CCL20 (R&D systems MAB360) at a concentration of 1:100 and goat anti-mouse IgG Alexa fluor 647 (ThermoFisher #A48289) at a concentration of 1:500.

Techniques: Expressing, Phospho-proteomics, Inhibition

Journal: Frontiers in Immunology

Article Title: Macrophage-derived CCL20 promotes abdominal aortic aneurysm progression via lymphocytes CCR6

doi: 10.3389/fimmu.2026.1780720

Figure Lengend Snippet: Macrophages Exhibit Altered Gene Expression and Secrete Abundant CCL20. (A) The volcano plot of DEGs of macrophages in the normal and AAA groups in GSE166676 and GSE226492 . (B) Bubble plot of GO BP enrichment results for DEGs. (C) Bubble plot of enriched KEGG pathways for DEGs. (D) GSEA results based on KEGG identified activated and suppressed pathways of macrophages in the normal and AAA groups. Categories with red dots (NES > 0) showed the activated pathways in AAA group, while categories with blue dots (NES < 0) showed the activated pathways in the normal group. (E) The mRNA expression of IL-1beta, IL-8, TNF-α in normal and AAA tissues from human. (F) The mRNA expression of ACC1, FASN, and PPAR-γ in human Normal and AAA groups. (G) Representative images of Oil red O staining of the normal (n=5) and AAA group (n=6). Scale bar:10, 25μm. (H) All up DEGs in the Cytokine-Cytokine receptor interaction pathway. (I) CCL20 expression in 11 cell types. (J) Representative images of IF co-localization of CCL20 (green) and macrophages (CD68, red). Nuclei were stained with DAPI (blue), scale bar, 50 μm. The synchronized fluctuations of CCL20 (green) and macrophages (CD68, red) curves indicated co-localization at these discrete sites. (K) CCL20 expression in 11 cell types between the normal group and AAA groups. (L) ELISA test of CCL20 in the normal group (n=79) and AAA group (n=80). (M) ROC curve comparing models for AAA diagnosis. The model achieved an area under the curve (AUC) of 0.7. (N) Representative images of IF of CCL20 (green) expression in the the normal (n=5) and AAA group (n=6). Nuclei were stained with DAPI (blue), scale bar, 50 μm. * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001.

Article Snippet: Serum CCL20 levels in AAA patients and healthy controls were quantified using a commercial ELISA kit (Proteintech, KE00149, China), following the manufacturer’s protocol.

Techniques: Gene Expression, Expressing, Staining, Enzyme-linked Immunosorbent Assay, Biomarker Discovery

Journal: Frontiers in Immunology

Article Title: Macrophage-derived CCL20 promotes abdominal aortic aneurysm progression via lymphocytes CCR6

doi: 10.3389/fimmu.2026.1780720

Figure Lengend Snippet: Macrophages Recruit Significant Numbers of Immune Cells through the CCL20-CCR6 Axis. (A) CCR6 expression in 11 cell types in GSE166676 and GSE226492 . (B) Representative images of IF co-localization of CCR6 (green) and B cells (CD19, red). Nuclei were stained with DAPI (blue), scale bar, 50μm. (C) Representative images of IF co-localization of CCR6 (green) and T cells (CD3, yellow). Nuclei were stained with DAPI (blue), scale bar, 50μm. The synchronized fluctuations of CCR6 (green)and T cells (CD3, yellow) curves indicated co-localization at these discrete sites. (D) CCR6 expression in 11 cell types. (E) CCR6 protein expression in human tissues between the normal group and the AAA group. (F, G) Representative images of IF of CCR6 (green) expression in the the normal (n=5) and AAA group (n=6). Nuclei were stained with DAPI (blue), scale bar, 50 μm. (H) CCR6 mRNA expression in GSE269845 and GSE183464 . (I) Cell Chat between macrophages and other cells in the CCL pathway signaling. (J) Representative images of IF of Macrophages (CD68, green), B cells (CD19, red), T cells (CD3, yellow) in the normal and AAA groups. Nuclei were stained with DAPI (blue), scale bar, 50 μm. (K) Flow cytometry of CD19 cells frequency in the LPS (100ng/ml) group and the LPS + CCL20 Antibody group (L) Flow cytometry of CD3 cells frequency in LPS (100ng/ml) group and the LPS + CCL20 Antibody group, * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: Serum CCL20 levels in AAA patients and healthy controls were quantified using a commercial ELISA kit (Proteintech, KE00149, China), following the manufacturer’s protocol.

Techniques: Expressing, Staining, Flow Cytometry

Journal: Frontiers in Immunology

Article Title: Macrophage-derived CCL20 promotes abdominal aortic aneurysm progression via lymphocytes CCR6

doi: 10.3389/fimmu.2026.1780720

Figure Lengend Snippet: Targeting the CCL20-CCR6 Axis Suppresses AAA Progression. (A) Schematic representation of study design, n=15 per group. (B) RT-qPCR of knockdown efficiencies for CCL20 and CCR6. (C) The AAA incidence. (D) The maximal external aortic diameter (mm). (E) Representative images of morphology of the whole aorta of all groups. (F) Representative vascular Doppler ultrasound images of the aorta of all groups. (N = 15). (G) Representative hematoxylin and eosin (HE), Elastica van Gieson (EVG), and elastin degradation grading, Masson trichrome staining. (H) Representative IF images of CCR6 (green), T cells (CD3, yellow), and B cells (CD19, red) in the aorta tissues of all groups. Nuclei were stained with DAPI (blue), scale bar, 20 μm. * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001.

Article Snippet: Serum CCL20 levels in AAA patients and healthy controls were quantified using a commercial ELISA kit (Proteintech, KE00149, China), following the manufacturer’s protocol.

Techniques: Quantitative RT-PCR, Knockdown, Staining

Journal: Frontiers in Immunology

Article Title: Macrophage-derived CCL20 promotes abdominal aortic aneurysm progression via lymphocytes CCR6

doi: 10.3389/fimmu.2026.1780720

Figure Lengend Snippet: The role of CCL20-CCR6 axis in AAA formation. Macrophage polarization was imbalanced, with enriched M1-like macrophages and elevated CCL20 secretion. CCL20 promoted the recruitment of CCR6+ immune cells (T and B cells) and AAA formation.

Article Snippet: Serum CCL20 levels in AAA patients and healthy controls were quantified using a commercial ELISA kit (Proteintech, KE00149, China), following the manufacturer’s protocol.

Techniques: