Journal: Nature

Article Title: Metabolic heterogeneity confers differences in melanoma metastatic potential

doi: 10.1038/s41586-019-1847-2

Figure Lengend Snippet: a-c , Western blot analysis of MCT1 ( a ), MCT2 ( b ), and MCT4 ( c ) in 3 efficiently (M405, M481, and UT10) and 4 inefficiently (M498, M528, M597 and M610) metastasizing xenografted melanomas. Positive and negative controls for MCT1 and MCT4 were HCC15 cells and MCT1 or MCT4 deficient HCC15 cells. MCF7 cells were a positive control for MCT2. The data are representative of 4 (MCT1), 2 (MCT2), and 2 (MCT4) experiments. d-e , Flow cytometric analysis of MCT1 surface expression in inefficiently ( d ) and efficiently ( e ) metastasizing melanomas. f , Enrichment of lactate m+3 normalized to 3PG m+3 in xenografted tumors after treatment with the MCT1 inhibitor, AZD3965, or DMSO control and [U- 13 C]lactate infusion (2 experiments per melanoma). The number of mice per treatment is indicated in each panel. g-i , Growth of subcutaneous tumors ( g ) in mice treated with AZD3965 (AZD) or DMSO control as well as the frequency of circulating melanoma cells in the blood (h) and metastatic disease burden based on bioluminescence imaging (i) . Data in h and i reflect 1 (UT10) or 2 experiments per melanoma, but only one representative experiment per melanoma is shown in g . j-k , Growth of subcutaneous tumors ( j ) and metastatic disease burden at endpoint by bioluminescence imaging ( k ) in mice transplanted with YUMM1.7, YUMM3.3, or YUMM5.2 mouse melanomas and treated with AZD3965 (AZD) or DMSO control (two experiments per melanoma). Data represent mean ± s.d. Statistical significance was assessed using t-tests ( f ), nparLD ( g ), mixed effects analysis ( j ) or Mann-Whitney tests ( h-i and k ).

Article Snippet: To analyze other markers, cells were stained with Alexa Fluor647-conjugated anti-human MCT1 (Bioss antibodies), Alexa Fluor488-conjugated anti-human CD147, PE-Vio770-conjugated anti-human CD98, Alexa Fluor700-conjugated anti-human β1-Integrin, FITC-conjugated anti-E-Cadherin (CD324) or PE/Cy7-conjugated anti-N-Cadherin (CD325).

Techniques: Western Blot, Positive Control, Expressing, Imaging, MANN-WHITNEY

Journal: Nature

Article Title: Metabolic heterogeneity confers differences in melanoma metastatic potential

doi: 10.1038/s41586-019-1847-2

Figure Lengend Snippet: a , Quantification of MCT1 relative to Actin bands from the western blot in comparing efficient versus inefficient metastasizers. b , Quantification of MCT4 relative to Actin bands from the western blot in comparing efficient versus inefficient metastasizers. c-d , Quantification of mean fluorescence intensities for MCT1 staining in the flow cytometry plots comparing efficient and inefficient metastasizers. HCC15 cells and MCT1 -deficient HCC15 cells were positive and negative controls ( c ). e-f , Immunofluorescence staining for MCT1 (green) in sections from subcutaneous tumors from inefficiently ( e , UM47) or efficiently ( f , M405) metastasizing melanomas. An adjacent section was stained with an antibody against S100b (a melanoma marker, green). Images are representative of three independent experiments per melanoma. g,h Immunofluorescence staining for MCT1 (green) in sections from subcutaneous tumors from inefficient ( g , M498, M610, and M597) and efficient ( h , M481, UT10, and M405) metastasizers. In each case, an adjacent section was stained with an antibody against S100b (a melanoma marker, green). Images are representative of results from two independent experiments per melanoma. i-j , While efficient metastasizers often exhibited cell surface staining ( j ), inefficient metastasizers typically exhibited diffuse cytoplasmic staining ( i ). Images are representative of results from two independent experiments per melanoma. All data represent mean ± s.d.. Statistical significance was assessed using Student’s t -tests ( a-b and d ).

Article Snippet: To analyze other markers, cells were stained with Alexa Fluor647-conjugated anti-human MCT1 (Bioss antibodies), Alexa Fluor488-conjugated anti-human CD147, PE-Vio770-conjugated anti-human CD98, Alexa Fluor700-conjugated anti-human β1-Integrin, FITC-conjugated anti-E-Cadherin (CD324) or PE/Cy7-conjugated anti-N-Cadherin (CD325).

Techniques: Western Blot, Fluorescence, Staining, Flow Cytometry, Immunofluorescence, Marker

Journal: Nature

Article Title: Metabolic heterogeneity confers differences in melanoma metastatic potential

doi: 10.1038/s41586-019-1847-2

Figure Lengend Snippet: a-c , Western blot analysis of MCT1 ( a ), MCT4 ( b ) and CD147 ( c ) in subcutaneous tumours versus metastatic liver (liv), kidney (kid), and pancreas (pan) nodules from NSG mice transplanted with three melanomas. d-g , Flow cytometry histograms of anti-MCT1 ( d, e ), or anti-CD147 ( f, g ) staining in melanoma cells from subcutaneous tumors or metastatic nodules from mice transplanted with M405 ( d, f ) or M481 ( e, g ) melanomas. h-o , Flow cytometry histograms and mean fluorescence intensities of anti-MCT1 ( h, i ), anti-CD147 ( j, k ), anti-CD98 ( l, m ), or anti-β1-Integrin ( n-o ) staining in melanoma cells from subcutaneous tumors treated with DMSO (control; black) or AZD3965 (MCT1 inhibitor; blue). The number of tumors/mice analyzed in each treatment is indicated within the bars in each panel (2 to 3 experiments). In all flow cytometric analyses, human melanoma cells were distinguished from mouse cells based on positivity for HLA-ABC and DsRed and negativity for mouse CD31/CD45/Ter119 staining (see and for gating strategy). p-u , Western blot analysis of IKKα ( p-r ) and IKKβ ( s-u ) in subcutaneous tumors from NSG mice treated with DMSO or AZD3965. All data represent mean ± s.d. Statistical significance was assessed with two-way ANOVAs ( i, k, m , and o ).

Article Snippet: To analyze other markers, cells were stained with Alexa Fluor647-conjugated anti-human MCT1 (Bioss antibodies), Alexa Fluor488-conjugated anti-human CD147, PE-Vio770-conjugated anti-human CD98, Alexa Fluor700-conjugated anti-human β1-Integrin, FITC-conjugated anti-E-Cadherin (CD324) or PE/Cy7-conjugated anti-N-Cadherin (CD325).

Techniques: Western Blot, Flow Cytometry, Staining, Fluorescence

Journal: Nature

Article Title: Metabolic heterogeneity confers differences in melanoma metastatic potential

doi: 10.1038/s41586-019-1847-2

Figure Lengend Snippet: a , Western blot analysis of MCT1 in subcutaneous tumors from mice xenografted with efficiently metastasizing melanomas transfected with scrambled control shRNA or shRNA1 or shRNA2 against MCT1 . HCC15 cells were used as a positive control and MCT1 -deficient HCC15 cells were used as a negative control (representative of 2 independent experiments). b , Western blot analysis of MCT4 in subcutaneous tumors from mice xenografted with efficiently metastasizing melanomas transfected with scrambled control shRNA or shRNA1 or shRNA2 against MCT4 . HCC15 cells were used as a positive control and MCT4 -deficient HCC15 cells were used as a negative control. c-e , Growth of subcutaneous tumors ( c ) in mice transplanted with melanomas transfected with scrambled control shRNA or shRNA1 or shRNA2 against MCT1 . The number of mice analyzed in each treatment is indicated in each panel (one experiment per melanoma). The frequency of circulating melanoma cells in the blood (d) and metastatic disease burden based on bioluminescence imaging (e) in the same mice. f , Western blot analysis of MCT1 in subcutaneous tumors transfected with scrambled control shRNA or shRNA1 or shRNA2 against MCT1 , with (OE) or without an shRNA-insensitive MCT1 cDNA. g-h , Growth of subcutaneous tumors ( g ) and metastatic disease burden at endpoint ( h ) in mice transplanted with melanomas transfected with scrambled control shRNA or shRNA1 or shRNA2 against MCT1 and an shRNA-insensitive MCT1 cDNA. i , Fold change in mean fluorescence intensity for CellRox DeepRed staining (ROS) in xenografted melanoma cells with scrambled control shRNA or shRNA1 or shRNA2 against MCT1 treated with AZD3965 or DMSO. All data represent mean ± s.d.. Statistical significance was assessed using nparLD followed by Benjamiani-Hochberg’s multiple comparisons adjustment ( c) , log2 one-way ANOVAs with Holm-Sidak’s multiple comparisons adjustment ( d-e and h ), mixed-effects analysis followed by Dunnett’s multiple comparisons adjustment ( g ), or log2 two-way ANOVA with Sidak’s multiple comparisons adjustment ( i ).

Article Snippet: To analyze other markers, cells were stained with Alexa Fluor647-conjugated anti-human MCT1 (Bioss antibodies), Alexa Fluor488-conjugated anti-human CD147, PE-Vio770-conjugated anti-human CD98, Alexa Fluor700-conjugated anti-human β1-Integrin, FITC-conjugated anti-E-Cadherin (CD324) or PE/Cy7-conjugated anti-N-Cadherin (CD325).

Techniques: Western Blot, Transfection, shRNA, Positive Control, Negative Control, Imaging, Fluorescence, Staining

Journal: Nature

Article Title: Metabolic heterogeneity confers differences in melanoma metastatic potential

doi: 10.1038/s41586-019-1847-2

Figure Lengend Snippet: a , Western blot analysis of MCT1 in wild-type parental YUMM1.7 melanoma cells as well as two lines from which MCT1 had been deleted using CRISPR. b-d , Growth of subcutaneous tumors ( b ), total metastatic disease burden at endpoint by bioluminescence imaging of visceral organs ( c ) and CellRox DeepRed staining of subcutaneous tumor cells ( d ). The number of mice analyzed in each treatment is indicated in each panel (one experiment; note that one mouse died in the KO#2 treatment before endpoint analysis). e , Western blot analysis of MCT1 in an inefficiently metastasizing melanoma (UM47) expressing MCT1 cDNA . f-g , Growth of subcutaneous tumors ( f ) and total metastatic disease burden at endpoint by bioluminescence imaging of visceral organs ( g ) from mice transplanted with these melanomas (one experiment; note that two mice died in the control treatment before endpoint analysis). All data represent mean ± s.d.. Statistical significance was assessed using one-way ANOVA followed by Dunnett’s multiple comparison adjustment ( b: day 25) or log2 one-way ANOVAs followed by Dunnett’s multiple comparisons adjustment ( c-d ), t-test ( f : day 90) or log2 t-test ( g ).

Article Snippet: To analyze other markers, cells were stained with Alexa Fluor647-conjugated anti-human MCT1 (Bioss antibodies), Alexa Fluor488-conjugated anti-human CD147, PE-Vio770-conjugated anti-human CD98, Alexa Fluor700-conjugated anti-human β1-Integrin, FITC-conjugated anti-E-Cadherin (CD324) or PE/Cy7-conjugated anti-N-Cadherin (CD325).

Techniques: Western Blot, CRISPR, Imaging, Staining, Expressing

Journal: Nature

Article Title: Metabolic heterogeneity confers differences in melanoma metastatic potential

doi: 10.1038/s41586-019-1847-2

Figure Lengend Snippet: a , Migration in transwell invasion assays of three melanomas treated with DMSO (control) or AZD3965 (MCT1 inhibitor), including representative images ( left ) and counts ( righ t) of the cells that migrated across the insert after 24 hours (one experiment with two to three replicate cultures per melanoma). b-c , Effect of acute treatment with AZD3965 (7 days) on the diameter of subcutaneous tumors, the frequency of circulating melanoma cells in the blood, and metastatic disease burden in mice with established M481 ( b ) or M405 ( c ) melanomas. Treatment was initiated when the subcutaneous tumors reached 2 cm in diameter (one experiment per melanoma with three mice per treatment). d , Efficiently metastasizing melanoma cells (M405) were subcutaneously transplanted into mice, allowed to spontaneously metastasize, then the primary tumors were resected to prolong survival and to allow the metastatic tumors that had formed prior to primary tumor resection to grow larger. Mice were treated with AZD3965 for the duration of the experiment, only prior to primary tumor resection, or only after primary tumor resection. e , Analysis of total metastatic disease burden at endpoint showing that metastatic disease burden was reduced when AZD3965 treatment was performed prior to primary tumor resection, during the time when melanoma cells were spontaneously metastasizing, but before metastatic tumors were established. The number of mice per treatment is shown in the panel (two independent experiments). All data represent mean ± s.d.. Statistical significance was assessed using two-way ANOVAs followed by Dunnett’s multiple comparison’s adjustment ( a ), t-tests ( b-c ) or Kruskal-Wallis test followed by Dunn’s multiple comparison’s adjustment ( e ).

Article Snippet: To analyze other markers, cells were stained with Alexa Fluor647-conjugated anti-human MCT1 (Bioss antibodies), Alexa Fluor488-conjugated anti-human CD147, PE-Vio770-conjugated anti-human CD98, Alexa Fluor700-conjugated anti-human β1-Integrin, FITC-conjugated anti-E-Cadherin (CD324) or PE/Cy7-conjugated anti-N-Cadherin (CD325).

Techniques: Migration

Journal: Nature

Article Title: Metabolic heterogeneity confers differences in melanoma metastatic potential

doi: 10.1038/s41586-019-1847-2

Figure Lengend Snippet: a-d , Kaplan-Meier overall survival curves of melanoma patients stratified based on MCT1 ( a ), MCT2 ( b ), MCT4 ( c ), and CD147 ( d ) expression level within tumor specimens. Data were from the SKCM cohort in TCGA ( https://portal.gdc.cancer.gov/projects/TCGA-SKCM ). Each panel compares the top third of patients with the highest expression levels versus the bottom third of patients with the lowest expression levels. Ticks represent censored values. e-f , Flow cytometry plots showing the gating strategies used to identify human melanoma cells in subcutaneous tumors ( e ) or the blood ( f ) of xenografted mice. Cells were gated on forward versus side scatter (FSC-A vs. SSC-A) to exclude red blood cells and clumps of cells. Human melanoma cells were selected by including cells that stained positively for DsRed (stably expressed in all melanoma lines) and HLA and excluding cells that stained positively for the mouse hematopoietic and endothelial markers CD45, CD31, or Ter119. The statistical significance of the differences in overall survival ( a-d ) were assessed using the Mantel-Cox log-rank test.

Article Snippet: To analyze other markers, cells were stained with Alexa Fluor647-conjugated anti-human MCT1 (Bioss antibodies), Alexa Fluor488-conjugated anti-human CD147, PE-Vio770-conjugated anti-human CD98, Alexa Fluor700-conjugated anti-human β1-Integrin, FITC-conjugated anti-E-Cadherin (CD324) or PE/Cy7-conjugated anti-N-Cadherin (CD325).

Techniques: Expressing, Flow Cytometry, Staining, Stable Transfection

Journal: Nature

Article Title: Metabolic heterogeneity confers differences in melanoma metastatic potential

doi: 10.1038/s41586-019-1847-2

Figure Lengend Snippet: a , Glutathione (GSH) to oxidized glutathione (GSSG) ratios in melanoma cells from mice treated with AZD3965 or DMSO (two independent experiments per melanoma). b , Quantitative analysis of NADPH and NADP+ in melanoma cells from mice treated with AZD3965 or DMSO (one or two experiments per melanoma). Liver cells were included as a control, with a high NADPH/NADP+ ratio. c , Expected isotope labelled species after [1,2- 13 C]glucose infusion. d , Glucose m+2 as a fraction of total plasma glucose in mice xenografted with efficiently metastasizing melanomas (M405, M481, and UT10), treated with DMSO or AZD3965, and infused with [1,2- 13 C]glucose. e , Glucose m+6 as a fraction of total plasma glucose in mice infused with [U- 13 C]glucose. The number of mice per treatment is indicated in each panel (two independent experiments). f-i , LC-MS measurement of the levels of glycolytic ( f, h ) and oxidative pentose phosphate pathway ( g, i ) metabolites in subcutaneous tumor cells from mice xenografted with melanomas treated with DMSO (control) or AZD3965 (MCT1 inhibitor) for 7 days. j , Flow cytometrically isolated MCT1 high or MCT1 −/low melanoma cells were subcutaneously transplanted into NSG mice, using 10 or 100 cells per injection. All injections formed tumors. Rate of growth of the tumors initiated with 10-cell injections. All data represent mean ± s.d.. Statistical significance was assessed using t-tests ( a ), repeated measures two-way ANOVAs ( b ), t-test ( e: 180 min), log2 two-way ANOVAs ( f and h ), log2 t-tests ( g: M405 and UT10), Mann-Whitney test ( g: M481 and i: M481), Welch’s t-tests ( i: M405 and UT10) or using nparLD test ( d and j ).

Article Snippet: To analyze other markers, cells were stained with Alexa Fluor647-conjugated anti-human MCT1 (Bioss antibodies), Alexa Fluor488-conjugated anti-human CD147, PE-Vio770-conjugated anti-human CD98, Alexa Fluor700-conjugated anti-human β1-Integrin, FITC-conjugated anti-E-Cadherin (CD324) or PE/Cy7-conjugated anti-N-Cadherin (CD325).

Techniques: Liquid Chromatography with Mass Spectroscopy, Isolation, Injection, MANN-WHITNEY

Journal: Nature

Article Title: Metabolic heterogeneity confers differences in melanoma metastatic potential

doi: 10.1038/s41586-019-1847-2

Figure Lengend Snippet: a-d , Flow cytometric analysis of anti-MCT1 staining in melanoma cells from subcutaneous tumors ( a,c ) or circulating melanoma cells ( b,d ) from the same mice xenografted with M405 ( a-b ) or M481 ( c-d ) ( and show the gating strategies to identify human melanoma cells; the data are representative of 3 experiments). e , Flow cytometrically isolated MCT1 high or MCT1 −/low melanoma cells were intravenously transplanted into NSG mice, using 100 or 1000 cells per injection. The panel shows the percentage of injections that formed metastatic tumors (1–2 experiments per melanoma). The number of mice analyzed per treatment is indicated in each panel. f , Metastatic disease burden in the visceral organs of mice that survived to endpoint after injection with 100 cells (M405 and M481) or 1000 cells (UT10) based on bioluminescence signal intensity. Data represent mean ± s.d. Statistical significance was assessed using multiple linear regression ( e ) or Mann-Whitney tests ( f ).

Article Snippet: To analyze other markers, cells were stained with Alexa Fluor647-conjugated anti-human MCT1 (Bioss antibodies), Alexa Fluor488-conjugated anti-human CD147, PE-Vio770-conjugated anti-human CD98, Alexa Fluor700-conjugated anti-human β1-Integrin, FITC-conjugated anti-E-Cadherin (CD324) or PE/Cy7-conjugated anti-N-Cadherin (CD325).

Techniques: Staining, Isolation, Injection, MANN-WHITNEY

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