Journal: Clinical and Vaccine Immunology

Article Title: Development and Characterization of Monoclonal Antibodies and Aptamers against Major Antigens of Mycobacterium avium subsp. paratuberculosis

doi: 10.1128/cvi.00022-07

Figure Lengend Snippet: FIG. 3. Evaluation of MAbs against whole-cell homogenates from several mycobacterial species. Immunoblot analysis shows that the reactivity of each MAb is observed with more than just M. avium subsp. paratuberculosis lysates. (A) Lanes: 1, M. silvaticum; 2, M. scrofula- ceum; 3, M. abscessus; 4, M. avium subsp. paratuberculosis K-10; 5, M. avium (strain TMC702); 6, M. bovis (strain 95-1315); 7, M phlei; 8, M. bovis BCG; 9, M. avium subsp. paratuberculosis ATCC 19698; 10, M. avium subsp. avium (strain TMC715); 11, M. avium subsp. paratuber- culosis (Linda); 12, M. intracellulare; 13, M. kansasii. (B) Lanes: 1, M. silvaticum; 2, M. scrofulaceum; 3, M. abscessus; 4, M. avium subsp. paratuberculosis K-10; 5, M. avium subsp. avium (strain TMC702); 6, M. bovis (strain 95-1315); 7, M phlei; 8, M. avium subsp. paratubercu- losis ATCC 19698; 9, M. avium subsp. avium (strain TMC715); 10, M. avium subsp. paratuberculosis (strain Linda); 11, M. intracellulare; 12, M. kansasii. Kilodalton size standards are indicated in the left margin, and the MAb used is indicated in the right margin.

Article Snippet: Mycobacterial isolates used in this study Isolate Organism Host Location Reference or source K-10 M. avium subsp. paratuberculosis Bovine Feces 20 19698 M. avium subsp. paratuberculosis Bovine Feces ATCC 19698 6100 M. avium subsp. paratuberculosis Human Ileum ATCC 43015 187 M. avium subsp. paratuberculosis Bovine Ileum Recent clinical isolate, NADC 523 M. avium subsp. paratuberculosis Bovine Ileum 803 M. avium subsp. paratuberculosis Bovine Ileum 6009 M. avium subsp. avium Bovine ATCC 35716 (TMC715) 6003 M. avium subsp. avium Chicken ATCC 35713 (TMC702) 6006 M. avium subsp. silvaticum Roe deer V1-72 6076 M. abscessus ATCC 19977 M. bovis Bovine Lymph node ATCC 19210 M. bovis BCG Pasteur Bovine Milk ATCC 35734 6081 M. kansasii Human ATCC 12478 6010 M. intracellulare Swine ATCC 35773 6083 M. phlei ATCC 11758 6077 M. scrofulaceum Human Lymph node a Underlined nucleotides represent restriction endonuclease cut sites.

Techniques: Western Blot

Journal: Clinical and Vaccine Immunology

Article Title: Development and Characterization of Monoclonal Antibodies and Aptamers against Major Antigens of Mycobacterium avium subsp. paratuberculosis

doi: 10.1128/cvi.00022-07

Figure Lengend Snippet: FIG. 6. Immunoblot (A) and dot blot (B) analyses of aptamers to MAP0105c. (A) The immunoblot containing mycobacterial whole-cell sonicated extracts was exposed to aptamer 94. Lane assignments: 1, M. silvaticum; 2, M. scrofulaceum; 3, M. abscessus; 4, M. avium subsp. para- tuberculosis K-10; 5, M. avium subsp. avium (strain TMC702); 6, M. bovis; 7, M. phlei; 8, M. bovis BCG; 9, M. avium subsp. paratuberculosis ATCC 19698; 10, M. avium subsp. avium (strain TMC715); 11, M. avium subsp. paratuberculosis (strain Linda); 12, M. intracellulare; 13, M. kansasii. Size standards are indicated in kilodaltons in the left margin. (B) The dot blot was exposed to the three aptamers, which are indicated above the blots. Proteins spotted to the membrane are indicated in the left margin, and the state of the proteins is indicated in the right margin. Abbreviations: MBP, MBP fused to the -peptide of LacZ; 218-4, an MBP fusion containing the N-terminal half of MAP0105c; 218-9, an MBP fusion containing the C-terminal half of MAP005c; K-10, a whole-cell lysate of M. avium subsp. paratuberculosis K-10; Avium, a whole-cell lysate of M. avium subsp. avium TMC715.

Article Snippet: Mycobacterial isolates used in this study Isolate Organism Host Location Reference or source K-10 M. avium subsp. paratuberculosis Bovine Feces 20 19698 M. avium subsp. paratuberculosis Bovine Feces ATCC 19698 6100 M. avium subsp. paratuberculosis Human Ileum ATCC 43015 187 M. avium subsp. paratuberculosis Bovine Ileum Recent clinical isolate, NADC 523 M. avium subsp. paratuberculosis Bovine Ileum 803 M. avium subsp. paratuberculosis Bovine Ileum 6009 M. avium subsp. avium Bovine ATCC 35716 (TMC715) 6003 M. avium subsp. avium Chicken ATCC 35713 (TMC702) 6006 M. avium subsp. silvaticum Roe deer V1-72 6076 M. abscessus ATCC 19977 M. bovis Bovine Lymph node ATCC 19210 M. bovis BCG Pasteur Bovine Milk ATCC 35734 6081 M. kansasii Human ATCC 12478 6010 M. intracellulare Swine ATCC 35773 6083 M. phlei ATCC 11758 6077 M. scrofulaceum Human Lymph node a Underlined nucleotides represent restriction endonuclease cut sites.

Techniques: Western Blot, Dot Blot, Sonication, Membrane

Journal: iScience

Article Title: Cellular and molecular profiling of collagenous gastritis implicates pathogenic CD4 + T cells

doi: 10.1016/j.isci.2025.114344

Figure Lengend Snippet: Single cell analysis of patients with collagenous gastritis (A) Workflow for the collection of biopsies from the duodenum and stomach of control and patients with CG. Black and gray arrows indicate which tissues (stomach and duodenum, respectively) were processed by which methods. X indicates the approximate location of all biopsies. (B) Controls (H1-6) and CG biopsies were processed as fresh, unsorted biopsies for gene expression (GEX) libraries; frozen biopsies were CD45-sorted and processed for GEX and T cell receptor (TCR) libraries. CG patient “A” had a pre- and post-steroid biopsy collected for CD45-sorting (“A”; “Apost”), and “D” had a second biopsy (“D2”). (C) UMAP of all patients/samples, both unsorted and CD45-sorted. (D) Violin plot of canonical cluster-defining gene expression across clusters from (C). (E) Cluster frequency per sample. See also and .

Article Snippet: CD45 (Intracellular Domain) (D9M8I) XP® Rabbit mAb #13917 , Cell Signaling Technology , 13917S; RRID: AB_2750898.

Techniques: Single-cell Analysis, Control, Gene Expression

Journal: iScience

Article Title: Cellular and molecular profiling of collagenous gastritis implicates pathogenic CD4 + T cells

doi: 10.1016/j.isci.2025.114344

Figure Lengend Snippet: Patients with Collagenous gastritis have an influx of highly activated T cells (A) CD4 + and CD8 + T cells were quantified by flow cytometry of the duodenum (top row) or stomach (bottom row). (B) Representative flow plots from (A). (C) CD4:CD8 ratio from (A). (D and E) PD-1 expression on duodenal CD8 + (D) or CD4 + T cells (E). (F and G) PD-1 expression on gastric CD8 + (F) or CD4 + T cells (G). (H) Representative CD8 and CD4 stains from tissue sections. All images at 20x objective. Scale bars 500 μm. (I) Quantification of (H) for intraepithelial (top row) or intraepithelial and stromal lymphocytes (bottom) from slides. Total cells per five high power fields (HPFs). (J) Quantification of PD-L1 and PD-1 stained tissue sections. (K) CD4: CD8 ratio from T cells captured by scRNA-seq (see D) in CD45-sorted samples. Significance assessed by the Wilcoxon rank test. (L) Heatmap of differential gene expression comparing control patients to each CG patient sample for CD8 and CD4 scRNA-seq clusters from CD45+ sorted cells. Genes involved in T cell activation/exhaustion are shown. Dot indicates significantly different expression between a patient sample and all control controls. Data are mean ± SEM; flow percentages were compared with a Student’s t test (flow) or Wilcoxon test (histology) followed by a Benjamini-Hochberg correction. Gene expression data compared with a Benjamini-Hochberg-corrected Wilcoxon rank-sum test. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001 See also and .

Article Snippet: CD45 (Intracellular Domain) (D9M8I) XP® Rabbit mAb #13917 , Cell Signaling Technology , 13917S; RRID: AB_2750898.

Techniques: Flow Cytometry, Expressing, Staining, Gene Expression, Control, Activation Assay

Journal: iScience

Article Title: Cellular and molecular profiling of collagenous gastritis implicates pathogenic CD4 + T cells

doi: 10.1016/j.isci.2025.114344

Figure Lengend Snippet: Resident memory CD8 T cells and expanded T cell clonotypes are reduced in patients with CG (A) T cells were sub-clustered from all patients/samples. T RM = Tissue resident memory; T EM = Effector memory; CTL = cytotoxic T lymphocyte. (B) Dot plot of cluster-defining genes for T cell sub-clusters. (C) Sub-cluster frequency per sample. (D) Distribution of T cells split by patient disease status. (E) Barplot of sub-clusters grouped by disease status of the patient. Mean ± SEM; adjusted p -value from BH-adjusted Wilcoxon rank test shown for p-adj <0.1. (F) UMAP from (A) of TCR expansion across all CD45-sorted samples; cells are colored by the TCR frequency. (G) Barplot of unique TCR clonotypes; expanded clonotypes are those with 2+ cells per unique TCR. Percentages above bars indicate the percentage of expanded clonotypes out of the total number of unique clonotypes. (H) Percentage of expanded T cells (2+ cells/clonotype) out of all T cells with TCRs sequenced. (I) Percentage of unique, expanded clonotypes out of all unique clonotypes; similar to (G), but grouped by disease status. Error bars are ±SEM; percentages were compared with a Wilcoxon rank test. See also .

Article Snippet: CD45 (Intracellular Domain) (D9M8I) XP® Rabbit mAb #13917 , Cell Signaling Technology , 13917S; RRID: AB_2750898.

Techniques:

Journal: iScience

Article Title: Cellular and molecular profiling of collagenous gastritis implicates pathogenic CD4 + T cells

doi: 10.1016/j.isci.2025.114344

Figure Lengend Snippet: Memory, Th17, and naive CD4 T cells are reduced, while cytotoxic CD4, Th1, Th2, and Treg populations are increased in patients with CG (A) CD4 T cells were sub-clustered from all patients/samples. (B) Dot plot of cluster-defining genes for CD4 T cell sub-clusters. (C) CD4 sub-cluster frequency per sample. (D) Distribution of T cells, colored by patient disease status. (E) Barplot of CD4 sub-clusters grouped by disease status of patient. Mean ± SEM; adjusted p -value from BH-adjusted Wilcoxon test shown for p-adj <0.1. (F) UMAP from (A) of CD4 clonal expansion across all CD45-sorted samples; cells are colored by the TCR clone frequency. (G) Barplot of unique CD4 TCR clonotypes; expanded clonotypes are those with 2+ cells per unique TCR. Percentages above bars indicate the percentage of expanded clonotypes out of the total number of unique clonotypes. “Patient H5” had fewer than 20 TCRs sequenced and was omitted from the plot. (H) Percentage of expanded T cells (2+ cells/clonotype) out of all T cells with TCRs sequenced (left); percentage of unique, expanded clonotypes out of all unique clonotypes, similar to (G), but grouped by disease status. Error bars are ±SEM; percentages were compared with a Wilcoxon rank test. N.S. not significant. See also .

Article Snippet: CD45 (Intracellular Domain) (D9M8I) XP® Rabbit mAb #13917 , Cell Signaling Technology , 13917S; RRID: AB_2750898.

Techniques:

Journal: iScience

Article Title: Cellular and molecular profiling of collagenous gastritis implicates pathogenic CD4 + T cells

doi: 10.1016/j.isci.2025.114344

Figure Lengend Snippet: Effects of steroid treatment on pre-existing TCR clonotypes and possible future therapies for CG (A) Hallmark Gene Set Enrichment Analysis of all cells from CD45-sorted patient “A”, comparing pre- and post-steroid gene expressions. (B) TCR clones sequenced from patient “A” show the distribution of clonotypes that were shared between time points (pre- and post-steroids) and expanded (2+ cells); expanded but not shared; shared but not expanded; or neither shared nor expanded (not). Percentage of total clone pool for each timepoint shown. (C) Alluvial plot from patient “A” showing unique TCR clones that were expanded (2+ cells) and shared between the two time points (pre- and post-steroids). Flow between bars indicates which sub-cluster the TCR clones were shared between. (D) The top four expanded TCR clones shared between pre- and post-steroids for patient “A” and their T cell sub-clusters of origin. Y axis indicates the total number of T cells captured in each shared clonotype. TCR chains are listed below every four clones. (E) Heatmap of scaled gene expression for T cell activation genes across T cell sub-clusters. Dot indicates significantly different expression between the CG and control samples for a particular gene/cluster. (F) Same as (E), but across CD4 sub-clusters. (G) Expression of IL5 and IL13 in the Th2 subcluster. (H) Differential gene expression analysis was performed across each T cell sub-cluster. Normalized JAK3 expression shown for T cell subclusters in which p-adj ≤0.05. Adjusted p -value and log2 fold-change (LFC) displayed. (I) Same as (H) but for CD4 sub-clusters in which p-adj ≤0.05. (J) Normalized ITGA4 expression shown for T cell subclusters in which p-adj ≤0.05. Adjusted p -value and log2 fold-change (LFC) displayed. (K) Same as (J), but for CD4 sub-clusters in which p-adj ≤0.05. (L) Normalized MADCAM1 (ligand for α4 integrin) on endothelial cells. Gene expression significance assessed by the Wilcoxon rank-sum test with a Benjamini-Hochberg correction. See also .

Article Snippet: CD45 (Intracellular Domain) (D9M8I) XP® Rabbit mAb #13917 , Cell Signaling Technology , 13917S; RRID: AB_2750898.

Techniques: Clone Assay, Gene Expression, Activation Assay, Expressing, Control

Journal: Integrative biology : quantitative biosciences from nano to macro

Article Title: Receptor for Hyaluronan Mediated Motility (RHAMM/HMMR) is a novel target for promoting subcutaneous adipogenesis

doi: 10.1039/c7ib00002b

Figure Lengend Snippet: A. Micro-CT images of 8-month female Wildtype, CD44-/- and RHAMM-/-. B. Quantification of total body mass. CD44-/- mice have significantly smaller body mass than wildtype counterparts while RHAMM-/- mice exhibit a significantly increased body mass. Values are the mean and S.E.M. n=3 mice/genotype. Asterisks indicate the difference between means is statistically significant and p values indicate the level of statistical significance using a two-tailed Student's T test.

Article Snippet: Polyclonal RHAMM anti-peptide antibodies were prepared against human synthetic peptides: KRFNDPSGCAPSPGAYD (PAb1), ELMKLRNKRETKMR (PAb2), KLKDENSQLKSEVSK (PAb 3) and CYRAPMECQESW (PAb4) (ProSci), chosen by analysis of hydrophilicity (Kyte-Doolittle), antigenic index (Jamesson-Wolf) and surface probability plots (Emini) for rat RHAMM (accession number {"type":"entrez-protein","attrs":{"text":"NP_037096","term_id":"47059017"}} NP_037096 ).

Techniques: Micro-CT, Two Tailed Test

Journal: Integrative biology : quantitative biosciences from nano to macro

Article Title: Receptor for Hyaluronan Mediated Motility (RHAMM/HMMR) is a novel target for promoting subcutaneous adipogenesis

doi: 10.1039/c7ib00002b

Figure Lengend Snippet: A. Micro CT image shows a typical optical section through the midsection of a mouse that was used to re-construct a 3-D image of subcutaneous fat (SubQ, extra-abdominal, arrow, lower image, SubQ quantified is outlined in red) and visceral fat (Visc, arrow) from which the values shown in the histogram were derived (as described in Experimental procedures). B. Micro CT analysis of total adipose tissue shows a significant increase in RHAMM-/- vs. wildtype mice and decrease in CD44-/- mice compared to wildtype counterparts. RHAMM-/- mice exhibit significantly increased subcutaneous (p=0.038) adipose tissue compared to wildtype mice while CD44-/- mice exhibit reduced SubQ and Visc. Asterisks indicate the difference between Wildtype and CD44-/- or between Wildtype and RHAMM-/- means are statistically significant and p values indicate the level of statistical significance using a two-tailed Student's T test.

Article Snippet: Polyclonal RHAMM anti-peptide antibodies were prepared against human synthetic peptides: KRFNDPSGCAPSPGAYD (PAb1), ELMKLRNKRETKMR (PAb2), KLKDENSQLKSEVSK (PAb 3) and CYRAPMECQESW (PAb4) (ProSci), chosen by analysis of hydrophilicity (Kyte-Doolittle), antigenic index (Jamesson-Wolf) and surface probability plots (Emini) for rat RHAMM (accession number {"type":"entrez-protein","attrs":{"text":"NP_037096","term_id":"47059017"}} NP_037096 ).

Techniques: Micro-CT, Construct, Derivative Assay, Two Tailed Test

Journal: Integrative biology : quantitative biosciences from nano to macro

Article Title: Receptor for Hyaluronan Mediated Motility (RHAMM/HMMR) is a novel target for promoting subcutaneous adipogenesis

doi: 10.1039/c7ib00002b

Figure Lengend Snippet: Paraffin processed histology sections of wildtype and RHAMM-/- skin were stained for adiponectin (brown stain) and counterstained with hematoxylin (blue stain). The adiponectin-positive brown stain was quantified using image analysis as described in Methods and results show that adiponectin protein levels are increased in RHAMM-/- subcutaneous adipose tissue. Values are the mean and S.E.M n=5 tissue sections. Asterisk indicates statistical significance between means, and p value indicates the level of statistical significance using a two-tailed Student's T test.

Article Snippet: Polyclonal RHAMM anti-peptide antibodies were prepared against human synthetic peptides: KRFNDPSGCAPSPGAYD (PAb1), ELMKLRNKRETKMR (PAb2), KLKDENSQLKSEVSK (PAb 3) and CYRAPMECQESW (PAb4) (ProSci), chosen by analysis of hydrophilicity (Kyte-Doolittle), antigenic index (Jamesson-Wolf) and surface probability plots (Emini) for rat RHAMM (accession number {"type":"entrez-protein","attrs":{"text":"NP_037096","term_id":"47059017"}} NP_037096 ).

Techniques: Staining, Two Tailed Test

Journal: Integrative biology : quantitative biosciences from nano to macro

Article Title: Receptor for Hyaluronan Mediated Motility (RHAMM/HMMR) is a novel target for promoting subcutaneous adipogenesis

doi: 10.1039/c7ib00002b

Figure Lengend Snippet: Polyclonal antibodies to RHAMM sequence were prepared as shown in the diagram. These were tested for their ability to promote adipogenesis (See Figure 5). Since PAb3 was the only antibody that promoted adipogenesis, synthetic peptides of varying lengths were prepared from the sequence used to generate PAb3 (NPI-0102, NPI0104). These were then tested for their effect on adipogenesis. The tubulin derived peptide, NPI-0109, which binds to the HA binding region of RHAMM and behaves as an HA fragment function blocking peptide mimic, was also tested for its ability to promote adipogenesis.

Article Snippet: Polyclonal RHAMM anti-peptide antibodies were prepared against human synthetic peptides: KRFNDPSGCAPSPGAYD (PAb1), ELMKLRNKRETKMR (PAb2), KLKDENSQLKSEVSK (PAb 3) and CYRAPMECQESW (PAb4) (ProSci), chosen by analysis of hydrophilicity (Kyte-Doolittle), antigenic index (Jamesson-Wolf) and surface probability plots (Emini) for rat RHAMM (accession number {"type":"entrez-protein","attrs":{"text":"NP_037096","term_id":"47059017"}} NP_037096 ).

Techniques: Sequencing, Derivative Assay, Binding Assay, Blocking Assay

Journal: Integrative biology : quantitative biosciences from nano to macro

Article Title: Receptor for Hyaluronan Mediated Motility (RHAMM/HMMR) is a novel target for promoting subcutaneous adipogenesis

doi: 10.1039/c7ib00002b

Figure Lengend Snippet: A. Bright field micrographs of rat bone marrow mesenchymal stem cells primed with glucocorticoids and then exposed to either medium alone (negative control), insulin (positive control, not shown), RHAMM antibody (PAb3) or a RHAMM peptide (NPI-0102 shown). Differentiation was detected by the presence of fat droplets, which were visualized by staining with the lipophilic dye Oil Red O. Both PAb-3 and RHAMM peptide promoted fat droplet accumulation. B. Oil Red O was extracted and quantified by measuring absorbance at 520 nm. Insulin, PAb-3 and RHAMM peptide (NPI-0104 shown) significantly stimulated (marked by asterisks) Oil Red O uptake above negative controls. C. RHAMM peptide (NPI-0102 shown) also stimulates adipocyte differentiation in human primary pre-adipocytes isolated from subcutaneous adipose depots as detected by oil red O uptake in lipid droplets. Adipogenesis stimulated by NPI-0102 is similar to the effect of a PPARγ agonist included as a positive control for these assays. Values are the means of n=5 replicates. P values indicate the level of statistical significance using a two-tailed Student's T test.

Article Snippet: Polyclonal RHAMM anti-peptide antibodies were prepared against human synthetic peptides: KRFNDPSGCAPSPGAYD (PAb1), ELMKLRNKRETKMR (PAb2), KLKDENSQLKSEVSK (PAb 3) and CYRAPMECQESW (PAb4) (ProSci), chosen by analysis of hydrophilicity (Kyte-Doolittle), antigenic index (Jamesson-Wolf) and surface probability plots (Emini) for rat RHAMM (accession number {"type":"entrez-protein","attrs":{"text":"NP_037096","term_id":"47059017"}} NP_037096 ).

Techniques: Negative Control, Positive Control, Staining, Isolation, Two Tailed Test

Journal: Integrative biology : quantitative biosciences from nano to macro

Article Title: Receptor for Hyaluronan Mediated Motility (RHAMM/HMMR) is a novel target for promoting subcutaneous adipogenesis

doi: 10.1039/c7ib00002b

Figure Lengend Snippet: The NPI-0104 peptide was injected under the right 4th nipple while the vehicle only control was injected under the nipple of the left 4th fat pad. Fat pads were photographed 7 days after injections. The areas of the treated and control fat pads were calculated from the photographs using image analysis. Results show that the NPI-0104 RHAMM function blocking peptide significantly increases mammary fat pad area (asterisks, p<0.05). Values are the means and S.E.M. of n=5 rats for each group. P value indicates the level of statistical significance using a two-tailed Student's T test.

Article Snippet: Polyclonal RHAMM anti-peptide antibodies were prepared against human synthetic peptides: KRFNDPSGCAPSPGAYD (PAb1), ELMKLRNKRETKMR (PAb2), KLKDENSQLKSEVSK (PAb 3) and CYRAPMECQESW (PAb4) (ProSci), chosen by analysis of hydrophilicity (Kyte-Doolittle), antigenic index (Jamesson-Wolf) and surface probability plots (Emini) for rat RHAMM (accession number {"type":"entrez-protein","attrs":{"text":"NP_037096","term_id":"47059017"}} NP_037096 ).

Techniques: Injection, Control, Blocking Assay, Two Tailed Test

Journal: Integrative biology : quantitative biosciences from nano to macro

Article Title: Receptor for Hyaluronan Mediated Motility (RHAMM/HMMR) is a novel target for promoting subcutaneous adipogenesis

doi: 10.1039/c7ib00002b

Figure Lengend Snippet: A. Micro-CT images were taken at the level of the 4th nipple on the 7th day after injection of either the NPI-0102 RHAMM function blocking peptide (right side of images) or vehicle alone (left side of images). B. Micro CT Images show the increase in subcutaneous fat in the area of the mammary fat pads resulting from RHAMM function blocking peptide injection. Left images are vehicle only and right images are peptide injected. All Images were taken 7 days after peptide injection. Following 3D reconstruction of the ROI shown in (A), the mass of the fat pads was calculated. Peptide treatment significantly increased fat pad mass. The values are the Means and S.E.M. n=5 rats, p value indicates the level of statistical significance using a two-tailed Student's T test.

Article Snippet: Polyclonal RHAMM anti-peptide antibodies were prepared against human synthetic peptides: KRFNDPSGCAPSPGAYD (PAb1), ELMKLRNKRETKMR (PAb2), KLKDENSQLKSEVSK (PAb 3) and CYRAPMECQESW (PAb4) (ProSci), chosen by analysis of hydrophilicity (Kyte-Doolittle), antigenic index (Jamesson-Wolf) and surface probability plots (Emini) for rat RHAMM (accession number {"type":"entrez-protein","attrs":{"text":"NP_037096","term_id":"47059017"}} NP_037096 ).

Techniques: Micro-CT, Injection, Blocking Assay, Two Tailed Test

Journal: Integrative biology : quantitative biosciences from nano to macro

Article Title: Receptor for Hyaluronan Mediated Motility (RHAMM/HMMR) is a novel target for promoting subcutaneous adipogenesis

doi: 10.1039/c7ib00002b

Figure Lengend Snippet: A. The NPI-0104 RHAMM function blocking peptide was injected subcutaneously into dorsal back skin and fat pad dimensions were measured at weekly intervals. Results show that fat pad retention was significantly above vehicle only controls for at least 2 weeks following a single injection of peptide (p<0.05). Values are the Mean and S.E.M. n=6 replicates/group. B. Values in A. are presented as the fold increase relative to vehicle controls. C. The NPI-0102 RHAMM function blocking peptide was injected into the right mammary fat pad and fat pad volume was quantified by micro CT. Mammary fat pads injected with peptide were significantly larger than those injected with vehicle only at 5 weeks. Values are the mean and S.E.M. n=7 animals/group. D. m-RNA was isolated from mammary fat pads in (B) and QPCR performed for perilipin 1 as described in methods. Results show that expression of perilipin 1 is strongly increased in peptide-treated mammary fat pads verifying that the tissue volume increase detected by micro CT in B results from increased adipogenesis. Values are the mean and S.E.M. n=4 replicates. P values indicate the level of statistical significance using a two-tailed Student's T test.

Article Snippet: Polyclonal RHAMM anti-peptide antibodies were prepared against human synthetic peptides: KRFNDPSGCAPSPGAYD (PAb1), ELMKLRNKRETKMR (PAb2), KLKDENSQLKSEVSK (PAb 3) and CYRAPMECQESW (PAb4) (ProSci), chosen by analysis of hydrophilicity (Kyte-Doolittle), antigenic index (Jamesson-Wolf) and surface probability plots (Emini) for rat RHAMM (accession number {"type":"entrez-protein","attrs":{"text":"NP_037096","term_id":"47059017"}} NP_037096 ).

Techniques: Blocking Assay, Injection, Micro-CT, Isolation, Expressing, Two Tailed Test

Journal: Integrative biology : quantitative biosciences from nano to macro

Article Title: Receptor for Hyaluronan Mediated Motility (RHAMM/HMMR) is a novel target for promoting subcutaneous adipogenesis

doi: 10.1039/c7ib00002b

Figure Lengend Snippet: A. mRNA transcriptome analyses of 10T1/2 mesenchymal progenitor cells stably overexpressing RHAMM show that master adipogenic transcription factors and target genes are suppressed by RHAMM expression, particularly PPARγ and adiponectin (ADIPOQ). B. A network of these RHAMM suppressed adipogenic genes was constructed using IPA software that identifies individual causal relationships curated from the literature. A connective tissue development and function network incorporated most of these genes including RHAMM (HMMR) MAP kinases ERK1,2 and p38, which regulate adipogenesis. C. NPI-0102 promotes mRNA expression of PPARγ and its target gene adiponectin. Values are the mean and S.E.M. n=4 replicates. P values indicate the level of statistical significance using a two-tailed Student's T test.

Article Snippet: Polyclonal RHAMM anti-peptide antibodies were prepared against human synthetic peptides: KRFNDPSGCAPSPGAYD (PAb1), ELMKLRNKRETKMR (PAb2), KLKDENSQLKSEVSK (PAb 3) and CYRAPMECQESW (PAb4) (ProSci), chosen by analysis of hydrophilicity (Kyte-Doolittle), antigenic index (Jamesson-Wolf) and surface probability plots (Emini) for rat RHAMM (accession number {"type":"entrez-protein","attrs":{"text":"NP_037096","term_id":"47059017"}} NP_037096 ).

Techniques: Stable Transfection, Expressing, Construct, Software, Two Tailed Test