mouse anti human ccl20  (R&D Systems)

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: Molecular Therapy. Nucleic Acids

Article Title: DNA vaccines targeting hemagglutinin from 18 subtypes of influenza A virus to antigen-presenting cells confer broad protection

doi: 10.1016/j.omtn.2025.102814

Figure Lengend Snippet: MIP1α-targeted HA vaccines induce cross-reactive antibodies and protection when delivered simultaneously, but not separately (A) Phylogenetic tree showing the classification of group 1 and group 2 HA. The scale bar represents % variation in amino acid sequences (influenza group 1 in blue, group 2 in orange). (B) Schematic of a homodimeric vaccine protein consisting of (i) a targeting unit (the CCR1/3/5 ligand Mip1α [CCL3]), (ii) a dimerization unit (a shortened hinge of exons h1 and h4 and the C H 3 region from human IgG3), and (iii) an antigenic unit (HA). (C) Secretion of vaccine proteins. HEK293E cells were transiently transfected with 1 μg of each indicated Mip1α-C H 3-HA plasmid. Secreted vaccine proteins were detected in the supernatant after 72 h by ELISA using an anti-C H 3 coating antibody and an anti-Mip1α detection antibody (mean ± SEM of technical triplicates). (D–F) BALB/c mice ( n = 6/group) were immunized once with 20 μg of Mip1α-C H 3-HA vaccines encoding a single HA subtype ( A). (D) IgG titers against H1–H18 were measured in sera pooled per group by ELISA at week 6 after immunization (mean, technical triplicates). (E) H1 PR8 -specific IgG was measured in sera at weeks 2 and 6 after vaccination (mean ± SEM, individual mice). (F) Six weeks after vaccination, mice were challenged with influenza PR8 virus (5L×D 50 ) and monitored for weight loss (mean ± SEM) in two separate experiments. Mice were euthanized upon reaching 20% weight loss. (G–J) BALB/c mice ( n = 10/group) were vaccinated twice at weeks 0 and 5 (arrows) with the indicated plasmid mixtures. (G–I) IgG antibodies against H1 Cal07 (G) and H1 PR8 (H) were measured in individual mice at weeks 3, 6, 8, and 10, and (I) H1 PR8 -specific IgG1 and IgG2a were measured at week 8 (mean ± SEM). (J) Five weeks after the boost, mice were challenged with PR8 influenza virus (5×LD 50 ) and monitored for weight loss. Survival curves represent mice reaching 20% weight loss as the humane endpoint. Statistical significance was calculated using: Kruskal-Wallis with Dunn’s multiple comparisons test in (E) and (G–I), two-way ANOVA with Dunnet’s multiple comparison in (F), and the Gehan-Breslow-Wilcoxon test against NaCl in (J). All groups were compared to NaCl at the final time point in (E), (G), and (H), or on day 8 in (F). Only significant values are shown.

Article Snippet: Secreted vaccine proteins were detected with a biotinylated mAb specific for C H 3 of hγ3 chains (HP6017, B3773, 1:3000, Sigma-Aldrich), biotinylated anti-MIP1α (MAB450, 1 μg/mL, R&D systems), or a biotinylated anti-A/B mAb (2H11, 1 μg/mL, a kind gift from Dr. Ellis L. Reinherz), followed by a streptavidin-alkaline phosphatase conjugate (1:3000, Calbiochem).

Techniques: Vaccines, Transfection, Plasmid Preparation, Enzyme-linked Immunosorbent Assay, Virus, Comparison