Journal: OncoImmunology

Article Title: An epitope-specific novel anti-EMMPRIN polyclonal antibody inhibits tumor progression

doi: 10.1080/2162402x.2015.1078056

Figure Lengend Snippet: Figure 1: EMMPRIN structure and peptide design, and specificity of 161-Ab: (A) Partial

Article Snippet: One strip was probed with the 1:1,000 diluted commercial rat anti-mouse EMMPRIN (MAB772, R&D systems) and then with the 1:5,000 diluted HRP-conjugated goat anti-rat IgG (112-035- D ow nl oa de d by [ U ni ve rs ity o f R eg in a] a t 0 6: 58 2 4 Fe br ua ry 2 01 6 062, Jackson) and served as positive control (P.C).

Techniques:

Journal: Cell & Bioscience

Article Title: Development of a novel human CD147 knock-in NSG mouse model to test SARS-CoV-2 viral infection

doi: 10.1186/s13578-022-00822-6

Figure Lengend Snippet: Schematic representation of genotyping primers used to confirm hCD147KI expression. A combination of 4 primers was used to screen mice (internal primers hCD147A and hCD147B) and confirm proper integration into the mouse CD147 allele (flanking primers BSGC and BSGD)

Article Snippet: To stain mouse CD147 (mCD147), polyclonal donkey anti-goat IgG secondary (Jackson ImmunoResearch; 705–035-003 [HRP]; 1:500) was used against primary goat anti-mouse CD147 antibody (R&D Systems; BAF772; 1:100).

Techniques: Expressing

Journal: Cell & Bioscience

Article Title: Development of a novel human CD147 knock-in NSG mouse model to test SARS-CoV-2 viral infection

doi: 10.1186/s13578-022-00822-6

Figure Lengend Snippet: H&E and IHC of human CD147 in hCD147KI het -NSG mice. Human CD147 was stained (HIM6; 1:500) in the ( A ) lung, ( B ) liver, ( C ) intestine, ( D ) heart, ( E ) brain, ( F ) spleen, ( G ) kidney, ( H ) testis, and ( I ) trachea in WT-NSG (top) and hCD147KI het -NSG (bottom) mice. Images were taken using an Olympus Inverted Light Microscope. Scale bar represents 100 µm

Article Snippet: To stain mouse CD147 (mCD147), polyclonal donkey anti-goat IgG secondary (Jackson ImmunoResearch; 705–035-003 [HRP]; 1:500) was used against primary goat anti-mouse CD147 antibody (R&D Systems; BAF772; 1:100).

Techniques: Staining, Light Microscopy

Journal: Cell & Bioscience

Article Title: Development of a novel human CD147 knock-in NSG mouse model to test SARS-CoV-2 viral infection

doi: 10.1186/s13578-022-00822-6

Figure Lengend Snippet: Flow cytometric analysis reveals proper dual-expression of both mCD147 and hCD147 in PBMCs and various organs. Representative contour plots of CD147 expression on WT-NSG (top) and hCD147KI het -NSG (bottom) cells from ( A ) PBMCs, ( B ) lung, ( C ) liver, and ( D ) spleen using antibodies targeting either mouse CD147 protein, human CD147 protein, or a combination of both antibodies (far right). Relative percentages are listed, and significant shifts highlighted in red. Gating was determined based on donkey anti-goat/mouse isotype IgG antibody background staining

Article Snippet: To stain mouse CD147 (mCD147), polyclonal donkey anti-goat IgG secondary (Jackson ImmunoResearch; 705–035-003 [HRP]; 1:500) was used against primary goat anti-mouse CD147 antibody (R&D Systems; BAF772; 1:100).

Techniques: Expressing, Staining

Journal: Cell & Bioscience

Article Title: Development of a novel human CD147 knock-in NSG mouse model to test SARS-CoV-2 viral infection

doi: 10.1186/s13578-022-00822-6

Figure Lengend Snippet: Diagram of proposed working hypothesis of CD147 in SARS-CoV-2 infection. (1) SARS-CoV-2 virions infect human cells via the canonical pathway where host Angiotensin-converting Enzyme 2 (ACE2) receptors bind to viral spike proteins (red) and facilitate viral entry and infection. (2) CD147 proteins, via binding to surface binding partners (e.g., E-selectin), facilitate cell–cell adhesion, membrane fusion, and intercellular transfer of SARS-CoV-2 virions. (3) Erythrocytes and platelets which strongly express CD147, bind SARS-CoV-2 virions, and increase thrombosis risk and other clinical manifestations of COVID-19

Article Snippet: To stain mouse CD147 (mCD147), polyclonal donkey anti-goat IgG secondary (Jackson ImmunoResearch; 705–035-003 [HRP]; 1:500) was used against primary goat anti-mouse CD147 antibody (R&D Systems; BAF772; 1:100).

Techniques: Infection, Binding Assay, Membrane

Journal: Scientific Reports

Article Title: MGAT1 and Complex N-Glycans Regulate ERK Signaling During Spermatogenesis

doi: 10.1038/s41598-018-20465-3

Figure Lengend Snippet: Gene expression changes in Mgat1 cKO germ cells. ( A ) Western blot analysis for basigin before and after endoglycosidase H treatment of germ cell lysates. Basigin from Mgat1 cKO germ cells was sensitive to Endo H digestion, consistent with a lack of complex N-glycans. ( B ) Hierarchical clustering of genes expressed in control versus Mgat1 cKO germ cell cDNA preparations from 22 dpp males, and from 3 replicate cDNA preparations of control versus Mgat1 cKO germ cell RNA pools obtained from 6 mice per genotype at 23 dpp. Red indicates high and green indicates low relative expression. ( C ) qRT-PCR validation of up-regulated genes in microarray data using cDNA from individual preparations used in microarray experiments. Bars represent mean ± SEM (gray, control; black Mgat1 cKO). Data are from two experiments performed in duplicate (n = 3 mice/group). * p < 0.05, ** p < 0.01. ( D ) qRT-PCR of down-regulated genes identified in microarray data performed on the same RNA samples used in B. Data are from two experiments performed in duplicate (n = 3 mice/group). * p < 0.05, ** p < 0.01. Both control and Mgat1 cKO transcripts were determined relative to Actb .

Article Snippet: CHO wild type (Pro-5) and Mgat1 null Lec1 CHO cells (Pro-5Lec1.3 C) were co-transfected with 3 μg mouse basigin plasmid cDNA (MC200670, Origene, Rockville, MD, USA) and 0.3 μg pSV2NEO in 6 well plates with Xtreme Gene HP DNA transfection reagent (#06 366 244 001, Sigma-Aldrich) at 1:3, DNA:reagent.

Techniques: Gene Expression, Western Blot, Control, Expressing, Quantitative RT-PCR, Biomarker Discovery, Microarray

Journal: Scientific Reports

Article Title: MGAT1 and Complex N-Glycans Regulate ERK Signaling During Spermatogenesis

doi: 10.1038/s41598-018-20465-3

Figure Lengend Snippet: The top network identified by IPA in 22 dpp Mgat1 cKO germ cells. ( A ) Molecules are represented as nodes (see legend). Nodes in red or green represent up-regulated or down-regulated genes, respectively. ( B ) Validation of Network 1 by qRT-PCR performed in duplicate on germ cell cDNA prepared from 3 mice in each group. Gray bars represent control and black bars represent Mgat1 cKO germ cells (mean ± SEM; * p < 0.05, ** p < 0.01).

Article Snippet: CHO wild type (Pro-5) and Mgat1 null Lec1 CHO cells (Pro-5Lec1.3 C) were co-transfected with 3 μg mouse basigin plasmid cDNA (MC200670, Origene, Rockville, MD, USA) and 0.3 μg pSV2NEO in 6 well plates with Xtreme Gene HP DNA transfection reagent (#06 366 244 001, Sigma-Aldrich) at 1:3, DNA:reagent.

Techniques: Biomarker Discovery, Quantitative RT-PCR, Control

Journal: Scientific Reports

Article Title: MGAT1 and Complex N-Glycans Regulate ERK Signaling During Spermatogenesis

doi: 10.1038/s41598-018-20465-3

Figure Lengend Snippet: Basigin signaling is reduced in Lec1 CHO cells, a model for Mgat1 cKO germ cells. Flow cytometry profiles of CHO wild type ( A ) and Lec1 mutant ( B ) CHO cells and basigin transfectants. Also shown is a complex N-glycan typical of CHO cells and the oligomannosyl N-glycan substrate of MGAT1 that accumulates on glycoproteins of Lec1 CHO cells. Blue square, GlcNAc; green circle, Man; yellow circle, Gal; red triangle, fucose; purple diamond, sialic acid. ( C ) CHO wild type, Lec1 and basigin transfectant lysates following 24 hr serum starvation and a 15 min incubation in serum-free medium were analyzed by western blotting using ERK Abs as shown. The gels from which these data were obtained are shown in Supplementary Fig. . ( D ) The pERK1/ERK1 and pERK2/ERK2 values for CHO, Lec1 and respective basigin transfectants (left panel), and the ratio of these values in basigin expressing versus non-basigin expressing CHO and Lec1 cells (right panel). Data are mean ± SEM from 9–13 gels of lysates run in 6 independent experiments. * p < 0.05 in ( D ) left panel is based on a one-tailed, unpaired Student’s t test with Welch’s correction; in ( E , left panel) significance is based on the non-parametric, two-tailed Wilcoxon matched-pairs signed rank test; in ( E ) right panel it is based on the unpaired, two-tailed Student’s t test with Welch’s correction. ** p < 0.01, *** p < 0.005 and **** p < 0.001.

Article Snippet: CHO wild type (Pro-5) and Mgat1 null Lec1 CHO cells (Pro-5Lec1.3 C) were co-transfected with 3 μg mouse basigin plasmid cDNA (MC200670, Origene, Rockville, MD, USA) and 0.3 μg pSV2NEO in 6 well plates with Xtreme Gene HP DNA transfection reagent (#06 366 244 001, Sigma-Aldrich) at 1:3, DNA:reagent.

Techniques: Flow Cytometry, Mutagenesis, Glycoproteomics, Transfection, Incubation, Western Blot, Expressing, One-tailed Test, Two Tailed Test

Journal: Frontiers in Immunology

Article Title: Serum EMMPRIN/CD147 promotes the lung pre-metastatic niche in a D2A1 mammary carcinoma mouse model

doi: 10.3389/fimmu.2025.1568578

Figure Lengend Snippet: EMMPRIN promotes primary tumor growth, and D2A1 cells do not metastasize into the lung in this model. (A) Female BALB/c mice were orthotopically injected with D2A1-WT cells, with D2A1-KD cells (2x10 5 cells each), or with no tumor cells (healthy). Alternatively, mice injected with D2A1-WT cells were i.p. treated with three injections of anti-EMMPRIN antibody (m161-pAb, 10 μg/ml/100μL, every 7 days), and healthy mice were i.p. injected with recombinant EMMPRIN (400 ng/ml/100μL, 6 times, every 4 days). After 28 days, mice were sacrificed, and their lungs were stained. (A) Representative images for the H&E staining (upper panel) or the immunohistochemical staining with the anti-mCherry antibody (lower panel). Bar size is 200 μm and 50 μm in the insets. The images demonstrate a denser and more congested lung structure in mice injected with the D2A1-WT cells or with the recombinant EMMPRIN, while mCherry remains unstained, indicating the lack of tumor cells in the lungs of all groups. (B) Tumor volumes or (C) tumor weights at the end of the experiment indicate that high levels of EMMPRIN promote tumor growth. (D) Wet lung weight was not changed, suggesting that metastatic mass was not added. Data are presented as mean ± SEM, and analyzed using one-way ANOVA followed by Bonferroni’s post hoc test, or the non-parametric two-tailed Mann-Whitney t test (n=14 for the D2A1-WT group, n=9 for the D2A1-KD group, n=9 for the healthy group, n=9 for the D2A1-WT + m161-pAb group, n=9 for the Healthy + rec. EMMRPIN group, in 2–3 biological repetitions).

Article Snippet: To simulate the effect of soluble EMMPRIN on the lung niche, healthy mice (Healthy) were intraperitoneally (i.p.) injected with 400 ng/ml/100μL of mouse recombinant EMMPRIN (R&D systems, Minneapolis, MN, USA) once every four days, starting on day 4, for a total of 6 injections.

Techniques: Injection, Recombinant, Staining, Immunohistochemical staining, Two Tailed Test, MANN-WHITNEY

Journal: Frontiers in Immunology

Article Title: Serum EMMPRIN/CD147 promotes the lung pre-metastatic niche in a D2A1 mammary carcinoma mouse model

doi: 10.3389/fimmu.2025.1568578

Figure Lengend Snippet: EMMPRIN serum levels correlate with tumor weight. Female BALB/c mice were orthotopically injected with D2A1-WT cells (n=13-14), with D2A1-KD cells (n=8-9), or with no tumor cells (healthy mice, n=9-10), or treated with the anti-EMMPRIN antibody (m161-pAb, n=9) or recombinant EMMPRIN protein (n=9) as described in the legend of Figure 1 . Mice were sacrificed at day 28 and EMMPRIN levels were determined by ELISA in the (A-C) serum, (D-F) lung lysates, and (G, H) tumor lysates. Serum, lung and tumor EMMPRIN levels were increased in the group injected with D2A1-WT cells relative to the group injected with D2A1-KD cells or to healthy mice, and these levels could be inhibited using the anti-EMMPRIN antibody or recapitulated by injecting recombinant EMMPRIN. Data are presented as mean ± SEM. Significance between three groups is analyzed using one-way ANOVA followed by Bonferroni’s post-hoc test, and two groups are analyzed using the non-parametric two-tailed Mann-Whitney t test. (I) Serum EMMPRIN levels were correlated with the primary tumor weight using the non-parametric Spearman correlation test. Blue and red dots indicate mice implanted with D2A1-WT or D2A1-KD cells, respectively.

Article Snippet: To simulate the effect of soluble EMMPRIN on the lung niche, healthy mice (Healthy) were intraperitoneally (i.p.) injected with 400 ng/ml/100μL of mouse recombinant EMMPRIN (R&D systems, Minneapolis, MN, USA) once every four days, starting on day 4, for a total of 6 injections.

Techniques: Injection, Recombinant, Enzyme-linked Immunosorbent Assay, Two Tailed Test, MANN-WHITNEY

Journal: Frontiers in Immunology

Article Title: Serum EMMPRIN/CD147 promotes the lung pre-metastatic niche in a D2A1 mammary carcinoma mouse model

doi: 10.3389/fimmu.2025.1568578

Figure Lengend Snippet: EMMPRIN promotes the secretion of VEGF, MMP-9 and TGFβ in the lung PMN. Mice were injected with the different tumor cells and treatments as described in the legend of Figure 1 . The levels of (A-C) VEGF, (D-F) MMP-9, and (G-I) TGFβ, were evaluated in the lung lysates and normalized to the total protein (n=12–13 for the D2A1-WT group, n=8–9 for the D2A1-KD group, n=9 for the healthy group, n=9 for the D2A1-WT + m161-pAb group, n=9 for the Healthy + rec. EMMRPIN group). Data are presented as mean ± SEM. Three groups were analyzed using one-way ANOVA followed by Bonferroni’s post-hoc test, and two groups were compared using the non-parametric two-tailed Mann-Whitney t test. The three cytokines were increased in the group injected with D2A1-WT cells relative to the group injected with D2A1-KD cells or to healthy mice, and the involvement of EMMPRIN in their regulation was demonstrated by the use of the anti-EMMPRIN antibody or by injecting recombinant EMMPRIN.

Article Snippet: To simulate the effect of soluble EMMPRIN on the lung niche, healthy mice (Healthy) were intraperitoneally (i.p.) injected with 400 ng/ml/100μL of mouse recombinant EMMPRIN (R&D systems, Minneapolis, MN, USA) once every four days, starting on day 4, for a total of 6 injections.

Techniques: Injection, Two Tailed Test, MANN-WHITNEY, Recombinant

Journal: Frontiers in Immunology

Article Title: Serum EMMPRIN/CD147 promotes the lung pre-metastatic niche in a D2A1 mammary carcinoma mouse model

doi: 10.3389/fimmu.2025.1568578

Figure Lengend Snippet: EMMPRIN promotes angiogenesis in the lung PMN. Mice were injected with the different tumor cells and treatments as described in the legend of Figure 1 . (A) Representative images of the endothelial cell marker CD31 staining in the lungs, and (C) their quantitation (n=9 for the D2A1-WT group, n=8 for the D2A1-KD group, n=7 for the healthy group, n=7 for the D2A1-WT + m161-pAb group, n=9 for the Healthy + rec. EMMRPIN group). Bar size is 100 μm. (B) Representative images of bEND3 endothelial cell migration length after their incubation for 18 h with serum-starvation media containing 5µg of lung lysates from the different experimental groups. (D) Quantitation of the migration length (n=14 for the D2A1-WT group, n=10 for the D2A1-KD group, n=8 for the healthy group, n=9 for the D2A1-WT + m161-pAb group, n=7 for the Healthy + rec. EMMRPIN group). Bar size is 150 μm. Data are presented as mean ± SEM. Three groups were analyzed using one-way ANOVA followed by Bonferroni’s post-hoc test, and two groups were compared using the non-parametric two-tailed Mann-Whitney t test. The increased CD31 staining of endothelial cells in the mice implanted with the D2A1-WT cells and their longer migration distance suggests that they proliferate and migrate, two properties necessary for angiogenesis, more than the endothelial cells in healthy mice or mice implanted with the D2A1-KD cells. The importance of EMMPRIN to this process is exemplified by the results from the addition of h161-pAb or the injection of recombinant EMMPRIN.

Article Snippet: To simulate the effect of soluble EMMPRIN on the lung niche, healthy mice (Healthy) were intraperitoneally (i.p.) injected with 400 ng/ml/100μL of mouse recombinant EMMPRIN (R&D systems, Minneapolis, MN, USA) once every four days, starting on day 4, for a total of 6 injections.

Techniques: Injection, Marker, Staining, Quantitation Assay, Migration, Incubation, Two Tailed Test, MANN-WHITNEY, Recombinant

Journal: Frontiers in Immunology

Article Title: Serum EMMPRIN/CD147 promotes the lung pre-metastatic niche in a D2A1 mammary carcinoma mouse model

doi: 10.3389/fimmu.2025.1568578

Figure Lengend Snippet: EMMPRIN promotes neutrophil infiltration and fibroblast activation in the lung PMN. Mice were injected with the different tumor cells and treatments as described in the legend of Figure 1 . (A) Representative images of the neutrophil marker Ly6G and (B) of the fibroblasts activation marker αSMA staining in the lungs. Bar size is 150 μM. (C-E) Quantification of neutrophil infiltration, and (F-H) of fibroblast activation in the different experimental groups (n=9–10 for the D2A1-WT group, n=8–9 for the D2A1-KD group, n=8–10 for the healthy group, n=6 for the D2A1-WT + m161-pAb group, n=6–8 for the Healthy + rec. EMMRPIN group). Data are presented as mean ± SEM. Three groups were analyzed using one-way ANOVA followed by Bonferroni’s post-hoc test, and two groups were compared using the non-parametric two-tailed Mann-Whitney t test. Infiltrated neutrophils and fibroblasts were activated more in the lungs of mice implanted with the D2A1-WT cells than healthy mice or mice implanted with D2A1-KD cells. EMMPRIN’s involvement in these processes is also demonstrated by the results of the administration of the anti-EMMPRIN antibody and the injection of recombinant EMMPRIN.

Article Snippet: To simulate the effect of soluble EMMPRIN on the lung niche, healthy mice (Healthy) were intraperitoneally (i.p.) injected with 400 ng/ml/100μL of mouse recombinant EMMPRIN (R&D systems, Minneapolis, MN, USA) once every four days, starting on day 4, for a total of 6 injections.

Techniques: Activation Assay, Injection, Marker, Staining, Two Tailed Test, MANN-WHITNEY, Recombinant

Journal: Frontiers in Immunology

Article Title: Serum EMMPRIN/CD147 promotes the lung pre-metastatic niche in a D2A1 mammary carcinoma mouse model

doi: 10.3389/fimmu.2025.1568578

Figure Lengend Snippet: EMMPRIN regulates the expression of interstitial collagens. Mice were injected with the different tumor cells and treatments as described in the legend of Figure 1 . (A) Representative images of lung sections from the different experimental groups that were stained with Masson’s Trichrome. Blue fibers indicate presence of collagen. Bar size is 150 μm. (B-D) Quantification of the collagen fibers stained by Masson’s Trichrome (n=9 for the D2A1-WT group, n=8 for the D2A1-KD group, n=9 for the healthy group, n=9 for the D2A1-WT + m161-pAb group, n=7 for the Healthy + rec. EMMRPIN group). Total RNA was extracted from the lungs, reverse transcribed and amplified for the determination of mRNA expression of (E-G) collagen 1A, (H-J) collagen 3A (K-M) LOX (n=11–12 for the D2A1-WT group, n=8–9 for the D2A1-KD group, n=9–10 for the healthy group, n=8–9 for the D2A1-WT + m161-pAb group, n=8–9 for the Healthy + rec. EMMRPIN group). Data are presented as mean ± SEM. Three groups were analyzed using one-way ANOVA followed by Bonferroni’s post-hoc test, and two groups were compared using the non-parametric two-tailed Mann-Whitney t test. The ECM in mice implanted with the D2A1-WT cells was denser with more collagen fibers, and the expression of interstitial collagens Col1A and Col3A, as well as the crosslinking enzyme LOX were higher than the healthy mice or mice implanted with D2A1-KD cells. Again, EMMPRIN’s involvement was demonstrated by the results of administration of the m161-pAb and the injection of recombinant EMMPRIN.

Article Snippet: To simulate the effect of soluble EMMPRIN on the lung niche, healthy mice (Healthy) were intraperitoneally (i.p.) injected with 400 ng/ml/100μL of mouse recombinant EMMPRIN (R&D systems, Minneapolis, MN, USA) once every four days, starting on day 4, for a total of 6 injections.

Techniques: Expressing, Injection, Staining, Reverse Transcription, Amplification, Two Tailed Test, MANN-WHITNEY, Recombinant

Journal: Frontiers in Immunology

Article Title: Serum EMMPRIN/CD147 promotes the lung pre-metastatic niche in a D2A1 mammary carcinoma mouse model

doi: 10.3389/fimmu.2025.1568578

Figure Lengend Snippet: EMMPRIN induces the STAT3 and ERK1/2 signaling pathways. The activation of different signaling pathways was assessed in lung lysates obtained from healthy mice or healthy mice injected with recombinant EMMPRIN from previous experiments, by determining the phosphorylation of key proteins in these pathways. The phosphorylation of (A) STAT3 and (B) ERK1/2 were evaluated using Intracellular DuoSet ELISA kits (n=7). Additionally, representative images of western blot analyses of two repetitions of the phosphorylation of (C) Akt 1/2/3, (D) ERK1/2, and (E) IκBα, and their quantification (n=5). Recombinant EMMPRIN increased the phosphorylation of STAT3 and ERK1/2, but not of Akt or IκBα, suggesting that the PI3K and the NF-κB pathways do not mediate EMMPRIN’s effect on the lung PMN.

Article Snippet: To simulate the effect of soluble EMMPRIN on the lung niche, healthy mice (Healthy) were intraperitoneally (i.p.) injected with 400 ng/ml/100μL of mouse recombinant EMMPRIN (R&D systems, Minneapolis, MN, USA) once every four days, starting on day 4, for a total of 6 injections.

Techniques: Protein-Protein interactions, Activation Assay, Injection, Recombinant, Phospho-proteomics, Enzyme-linked Immunosorbent Assay, Western Blot

Journal: BMC cancer

Article Title: CD73 complexes with emmprin to regulate MMP-2 production from co-cultured sarcoma cells and fibroblasts.

doi: 10.1186/s12885-019-6127-x

Figure Lengend Snippet: Fig. 1 MS analysis of emmprin complexes identified CD73 and CD99. Proteins forming complexes with emmprin were identified from cancer cells alone or from co-cultures of cancer cells and fibroblasts, and were analyzed by immunoprecipitation, cross-linking, and mass spectrometric (MS) protein identification. A total of 548 protein molecules were identified using MS. Overlap between proteins identified in different conditions (tumor cell only or three molecular weight regions under co-culture conditions #1–3) is shown. CD73 and CD99 identified in the overlap of all three co- culture conditions were selected for investigation of their effect on regulation of MMP-2 production

Article Snippet: SDS-PAGE and immunoblotting were performed using 4–15% gradient gel (Bio-Rad, Hercules, CA) and antibodies against emmprin (mouse monoclonal, R&D System, Flanders, NJ), anti-CD73 (rabbit monoclonal, Cell Signaling, Danvers, MA), MMP-2 (monoclonal antibody, Daiichi Fine Chemical, Toyama, Japan) and MT1-MMP (Millipore, Bedford, MA).

Techniques: Immunoprecipitation, Molecular Weight, Co-Culture Assay

Journal: BMC cancer

Article Title: CD73 complexes with emmprin to regulate MMP-2 production from co-cultured sarcoma cells and fibroblasts.

doi: 10.1186/s12885-019-6127-x

Figure Lengend Snippet: Fig. 2 CD99 and CD73 form complexes with emmprin. a Complex formation between emmprin and CD99 was identified by immunoprecipitation and immunoblotting, performed in co-cultures of tumor cells and fibroblasts. Arrow indicates emmprin-CD99 complex. b Complex formation between emmprin and CD73 was identified by immunoprecipitation and immunoblotting, performed in co-cultures of tumor cells and fibroblasts. Arrow indicates emmprin-CD73 complex.

Article Snippet: SDS-PAGE and immunoblotting were performed using 4–15% gradient gel (Bio-Rad, Hercules, CA) and antibodies against emmprin (mouse monoclonal, R&D System, Flanders, NJ), anti-CD73 (rabbit monoclonal, Cell Signaling, Danvers, MA), MMP-2 (monoclonal antibody, Daiichi Fine Chemical, Toyama, Japan) and MT1-MMP (Millipore, Bedford, MA).

Techniques: Immunoprecipitation, Western Blot

Journal: BMC cancer

Article Title: CD73 complexes with emmprin to regulate MMP-2 production from co-cultured sarcoma cells and fibroblasts.

doi: 10.1186/s12885-019-6127-x

Figure Lengend Snippet: Fig. 5 Colocalization of emmprin/CD73 detected by immunofluorescent staining and in situ proximity ligation assay. Cytoplasmic CD73 (green) expression was observed in fibroblasts, tumor cells and co-culture cells. Membranous emmprin expression (red) was observed in tumor cells and co-cultured cells. Nuclei were stained with DAPI (blue). CD73 and emmprin were colocalized in tumor cells (b) and co-cultured cells (c). The green arrow points to fibroblasts expresssing green florescence (CD73); The yellow arrow points to tumor cells expressed yellow florescence (emmprin and CD73) (c). CD73 siRNA treatment causes downregulation of CD73 cytoplasmic expression, although, membranous emmprin expression was retained in co-cultured cells (d). The fluorescent red spots observed using in situ proximity ligation assay (PLA), indicating protein- protein colocalization in cells, confirmed the interaction between CD73 and emmprin. The detected dimers (emmprin/CD73) are represented as red dots in co-cultured cells (g). In cells transfected with CD73 siRNA prior to in situ PLA for emmmprin-CD73 interaction, CD73 siRNA treatment caused downregulation of red dots (h). Immunofluorescent staining, IF (a-d); In situ proximity ligation assay, PLA (e-h); fibroblast only (a, e); tumor cell only (b, f); co-culture (c-d, g-h)

Article Snippet: SDS-PAGE and immunoblotting were performed using 4–15% gradient gel (Bio-Rad, Hercules, CA) and antibodies against emmprin (mouse monoclonal, R&D System, Flanders, NJ), anti-CD73 (rabbit monoclonal, Cell Signaling, Danvers, MA), MMP-2 (monoclonal antibody, Daiichi Fine Chemical, Toyama, Japan) and MT1-MMP (Millipore, Bedford, MA).

Techniques: Staining, In Situ, Proximity Ligation Assay, Expressing, Co-Culture Assay, Cell Culture, Transfection

Journal: BMC cancer

Article Title: CD73 complexes with emmprin to regulate MMP-2 production from co-cultured sarcoma cells and fibroblasts.

doi: 10.1186/s12885-019-6127-x

Figure Lengend Snippet: Fig. 7 Expression of CD73 and emmprin in epithelioid sarcoma. The hematoxylin and eosin (H&E) section shows proliferation of severely atypical polygonal cells with enlarged hyperchromatic nuclei, forming irregular nests, accompanied by fibroblastic cells and fibrous stroma (a). Immunohistochemical (b-c) and fluorescent immunohistochemical (d-f) expression of CD73 and emmprin in epithelioid sarcoma specimen was examined. Both tumor cells and surrounding stromal cells were positive for CD73 (b). Membranous emmprin expression was observed only in tumor cells (c). Cytoplasmic CD73 (green) expression was observed in fibroblasts and in tumor cells (e). Membranous emmprin expression (red) was observed in tumor cells (f). Marger of figures (e) and (f). The green arrow, indicates fibroblasts expressing green florescence (CD73); The Yellow arrow, indicates tumor cells expressing yellow florescence (emmprin and CD73) (d). CD73 and emmprin were colocalized in tumor cells (e). Nuclei were stained with DAPI (blue). Expression of CD73 and emmprin was examined immunohistochemically in a total of ten tumors. All tumors show similar expression pattern (Additional file 6: Table S1)

Article Snippet: SDS-PAGE and immunoblotting were performed using 4–15% gradient gel (Bio-Rad, Hercules, CA) and antibodies against emmprin (mouse monoclonal, R&D System, Flanders, NJ), anti-CD73 (rabbit monoclonal, Cell Signaling, Danvers, MA), MMP-2 (monoclonal antibody, Daiichi Fine Chemical, Toyama, Japan) and MT1-MMP (Millipore, Bedford, MA).

Techniques: Expressing, Immunohistochemical staining, Staining

Journal: BMC cancer

Article Title: CD73 complexes with emmprin to regulate MMP-2 production from co-cultured sarcoma cells and fibroblasts.

doi: 10.1186/s12885-019-6127-x

Figure Lengend Snippet: Fig. 8 A pictorial representation of the interaction between emmprin on tumor cells and CD73 on fibroblasts. Emmprin mainly exists on tumor cells, and CD73 exists both on tumor cells and fibroblasts. Emmprin forms a complex with CD73, and regulates MMP-2 production in co-cultures of tumor cells and fibroblasts. Pro- MMP-2 produced by fibroblasts is probably activated by MT1-MMP expressed on tumor cells

Article Snippet: SDS-PAGE and immunoblotting were performed using 4–15% gradient gel (Bio-Rad, Hercules, CA) and antibodies against emmprin (mouse monoclonal, R&D System, Flanders, NJ), anti-CD73 (rabbit monoclonal, Cell Signaling, Danvers, MA), MMP-2 (monoclonal antibody, Daiichi Fine Chemical, Toyama, Japan) and MT1-MMP (Millipore, Bedford, MA).

Techniques: Produced