Journal: Virology

Article Title: Human cytomegalovirus-infected cells release extracellular vesicles that carry viral surface proteins

doi: 10.1016/j.virol.2018.08.008

Figure Lengend Snippet: (A) stain-free blot representing the spectrum of proteins obtained for HCMV and EV fractions after separation of AD169 HCMV pre-paration using a iodixanol step-gradient centrifugation. (B) Detection of EV-associated proteins CD63 (~60 kDa), Rab27A (~30 kDa), and calnexin (~25 kDa) proteins in both fractions. (C) Detection of HCMV capsid protein MCP (~150 kDa) and HCMV tegument protein, pp150 (~160 kDa) in HCMV and EV fractions after separation of AD169 viral preparation using a iodixanol step-gradient centrifugation.

Article Snippet: 10 μg of proteins were loaded on a 4–20% precast polyacrylamide gel (Bio-Rad Laboratories, Hercules, CA) and separated by SDS-PAGE, then transferred to PVDF membranes and probed with anti-CD63 (1 μg/ml, Thermo Fisher Scientific, Waltham, MA), anti-Calnexin (1 μg/ml, Thermo Fisher Scientific, Waltham, MA), anti-Rab27A (1 μg/ml, Thermo Fisher Scientific, Waltham, MA), anti-HCMV capsid MCP (2 μg/ml), and anti-HCMV tegument pp150 (2 μg/ml) monoclonal primary anti-mouse monoclonal antibodies (clones 28–4 and 36–14) and then goat peroxidase-conjugated anti-mouse IgG secondary anti-body (Bio-Rad Laboratories, Hercules, CA).

Techniques: Staining, Gradient Centrifugation

Journal: Anticancer research

Article Title: Epithelial–Mesenchymal Expression Phenotype of Primary Melanoma and Matched Metastases and Relationship with Overall Survival

doi: 10.21873/anticanres.11243

Figure Lengend Snippet: Cadherin expression in 42 melanoma primary lesions and their corresponding metastases

Article Snippet: This study was approved by the Dartmouth College Committee for the Protection of Human Subjects/Institutional Review Board (CPHS# 23388). table ft1 table-wrap mode="anchored" t5 caption a7 Cases, n Gender Male 28 Female 14 Primary site Head/neck 6 Trunk 7 Upper extremity 15 Lower extremity 14 Ulceration Present 26 Absent 16 Histological type Superficial spreading 10 Nodular 9 Acral lentiginous 4 NOS 19 Clinical stage at diagnosis I 2 IB/II 1 II 11 II/III 4 III 21 IV 2 Could not be determined 1 Open in a separate window NOS: Not otherwise specified. caption a8 Summary of tumor cases and histology Immunohistochemistry Immunohistochemical stains were performed on 4-μm paraffin sections with the following primary antibodies: E-cadherin (clone 36, dilution 1:400; BD Transduction Laboratories, San Jose,CA, USA) and N-cadherin (clone 3B9, dilution 1:150;Invitrogen Life Technologies, Waltham, MA, USA).

Techniques: Expressing, Staining

Journal: Anticancer research

Article Title: Epithelial–Mesenchymal Expression Phenotype of Primary Melanoma and Matched Metastases and Relationship with Overall Survival

doi: 10.21873/anticanres.11243

Figure Lengend Snippet: Representative immunohistochemical expression patterns of E-cadherin (a-f) and N-cadherin (g-l) in primary and metastatic melanomas. Case 1 (a, d, g, j) had strong membranous E-cadherin staining in the primary lesion (a), but loss of membranous E-cadherin staining with an increase of its nuclear staining in the matched metastatic lesion (d); N-cadherin expression was negative in both primary and metastatic lesions of this case (g and j). Case 2 (b, e, h, k) had strong membranous E-cadherin staining and moderate membranous N-cadherin staining in the primary lesion (b and h); membranous E-cadherin staining was weak and N-cadherin staining negative in its matched metastatic lesion (e and k). Case 3 (c, f, i, l) had focal weak membranous E-cadherin staining and weak membranous N-cadherin staining in the primary lesion (c and i); membranous E-cadherin staining was focal and moderate and membranous N-cadherin staining was strong in the corresponding metastatic lesion (f and l). Original magnification, ×400.

Article Snippet: This study was approved by the Dartmouth College Committee for the Protection of Human Subjects/Institutional Review Board (CPHS# 23388). table ft1 table-wrap mode="anchored" t5 caption a7 Cases, n Gender Male 28 Female 14 Primary site Head/neck 6 Trunk 7 Upper extremity 15 Lower extremity 14 Ulceration Present 26 Absent 16 Histological type Superficial spreading 10 Nodular 9 Acral lentiginous 4 NOS 19 Clinical stage at diagnosis I 2 IB/II 1 II 11 II/III 4 III 21 IV 2 Could not be determined 1 Open in a separate window NOS: Not otherwise specified. caption a8 Summary of tumor cases and histology Immunohistochemistry Immunohistochemical stains were performed on 4-μm paraffin sections with the following primary antibodies: E-cadherin (clone 36, dilution 1:400; BD Transduction Laboratories, San Jose,CA, USA) and N-cadherin (clone 3B9, dilution 1:150;Invitrogen Life Technologies, Waltham, MA, USA).

Techniques: Immunohistochemical staining, Expressing, Staining

Journal: Anticancer research

Article Title: Epithelial–Mesenchymal Expression Phenotype of Primary Melanoma and Matched Metastases and Relationship with Overall Survival

doi: 10.21873/anticanres.11243

Figure Lengend Snippet: Comparison of cadherin expression in primary versus metastatic melanomas

Article Snippet: This study was approved by the Dartmouth College Committee for the Protection of Human Subjects/Institutional Review Board (CPHS# 23388). table ft1 table-wrap mode="anchored" t5 caption a7 Cases, n Gender Male 28 Female 14 Primary site Head/neck 6 Trunk 7 Upper extremity 15 Lower extremity 14 Ulceration Present 26 Absent 16 Histological type Superficial spreading 10 Nodular 9 Acral lentiginous 4 NOS 19 Clinical stage at diagnosis I 2 IB/II 1 II 11 II/III 4 III 21 IV 2 Could not be determined 1 Open in a separate window NOS: Not otherwise specified. caption a8 Summary of tumor cases and histology Immunohistochemistry Immunohistochemical stains were performed on 4-μm paraffin sections with the following primary antibodies: E-cadherin (clone 36, dilution 1:400; BD Transduction Laboratories, San Jose,CA, USA) and N-cadherin (clone 3B9, dilution 1:150;Invitrogen Life Technologies, Waltham, MA, USA).

Techniques: Expressing

Journal: Anticancer research

Article Title: Epithelial–Mesenchymal Expression Phenotype of Primary Melanoma and Matched Metastases and Relationship with Overall Survival

doi: 10.21873/anticanres.11243

Figure Lengend Snippet: Co-expression patterns of E-cadherin and N-cadherin

Article Snippet: This study was approved by the Dartmouth College Committee for the Protection of Human Subjects/Institutional Review Board (CPHS# 23388). table ft1 table-wrap mode="anchored" t5 caption a7 Cases, n Gender Male 28 Female 14 Primary site Head/neck 6 Trunk 7 Upper extremity 15 Lower extremity 14 Ulceration Present 26 Absent 16 Histological type Superficial spreading 10 Nodular 9 Acral lentiginous 4 NOS 19 Clinical stage at diagnosis I 2 IB/II 1 II 11 II/III 4 III 21 IV 2 Could not be determined 1 Open in a separate window NOS: Not otherwise specified. caption a8 Summary of tumor cases and histology Immunohistochemistry Immunohistochemical stains were performed on 4-μm paraffin sections with the following primary antibodies: E-cadherin (clone 36, dilution 1:400; BD Transduction Laboratories, San Jose,CA, USA) and N-cadherin (clone 3B9, dilution 1:150;Invitrogen Life Technologies, Waltham, MA, USA).

Techniques: Expressing

Journal: Anticancer research

Article Title: Epithelial–Mesenchymal Expression Phenotype of Primary Melanoma and Matched Metastases and Relationship with Overall Survival

doi: 10.21873/anticanres.11243

Figure Lengend Snippet: Multivariate analysis of the association of E-cadherin and N-cadherin expression on overall survival

Article Snippet: This study was approved by the Dartmouth College Committee for the Protection of Human Subjects/Institutional Review Board (CPHS# 23388). table ft1 table-wrap mode="anchored" t5 caption a7 Cases, n Gender Male 28 Female 14 Primary site Head/neck 6 Trunk 7 Upper extremity 15 Lower extremity 14 Ulceration Present 26 Absent 16 Histological type Superficial spreading 10 Nodular 9 Acral lentiginous 4 NOS 19 Clinical stage at diagnosis I 2 IB/II 1 II 11 II/III 4 III 21 IV 2 Could not be determined 1 Open in a separate window NOS: Not otherwise specified. caption a8 Summary of tumor cases and histology Immunohistochemistry Immunohistochemical stains were performed on 4-μm paraffin sections with the following primary antibodies: E-cadherin (clone 36, dilution 1:400; BD Transduction Laboratories, San Jose,CA, USA) and N-cadherin (clone 3B9, dilution 1:150;Invitrogen Life Technologies, Waltham, MA, USA).

Techniques: Expressing

Journal: Oncotarget

Article Title: The Hippo transducer TAZ promotes cell proliferation and tumor formation of glioblastoma cells through EGFR pathway

doi: 10.18632/oncotarget.9199

Figure Lengend Snippet: ( A, B ) The effects of TAZ on the colony formation in TAZ-knockdown U87 cells and TAZ-overexpressing U87 cells. ( C ) The tumor growth curve of TAZ-knockdown U87 cells injected into immunodeficient mice. ( D ) The tumor growth curve of TAZ-overexpressing U87 cells injected into immunodeficient mice. ( E ) Immunohistochemical staining for Ki67 in tumor tissues, shTAZ: shRNA for TAZ; shCtr: shRNA for control. Values are shown as the mean ± SD. * p < 0.05, ** p < 0.01. All p values are based on analysis control versus treatment.

Article Snippet: For immunohistochemistry, after quenching of endogenous peroxidase activity and blocking with normal goat serum, sections were incubated sequentially with Ki67 primary antibodies (1:100, clone 550609, BD pharmingen), biotinylated goat anti-mouse IgG, and the ABC reagent (Vector Laboratories).

Techniques: Injection, Immunohistochemical staining, Staining, shRNA

Journal: Molecular Cancer

Article Title: Radiation-induced caveolin-1 associated EGFR internalization is linked with nuclear EGFR transport and activation of DNA-PK

doi: 10.1186/1476-4598-7-69

Figure Lengend Snippet: Radiation induced caveolin-1 linked EGFR internalization is triggered by src kinase . Confluent A549 cells were irradiated with 4 Gy (A). At the time points given, src protein was immune-precipitated from cytoplasmic protein fraction. Proteins were separated by SDS-PAGE and after blotting the protein amounts of src, caveolin1 and EGFR were quantified. The same procedure was applied after cell stimulation with EGF (B). Phosphorylation of EGFR at the tyrosine No. 845, 992 and 1173 was quantified with help of specific antibodies following a standard western procedure. Equal protein amounts for immune-precipitation were documented by showing actin expression within aliquots of input proteins. (C). Protein expression was quantified by densitometry and shown as fold-induction relative to untreated control. The mean protein expression derived from three experiments was given below each band.

Article Snippet: The primary antibodies were diluted as follows: anti-EGFR (BD Transduction Laboratories, clone 13) 1:1000; anti EGFR pY845 (nanotools, clone 12A3) 1:1000; anti-EGFR pY992 (abcam, polyclonal) 1:500); anti-EGFR pY1173 (Cell signaling, clone 53A5) 1:1000; anti-phosphotyrosin (Santa Cruz, clone PY20) 1:500); anti-src (Santa Cruz, clone H-12) 1:1000; anti-src Y416 (cell signaling, polyclonal), 1:1000; anti-caveolin-1 (BD Transduction Laboratories, clone 2297) 1:1000; anti-caveolin pY14 (BD Transduction Laboratories, clone 56) 1:1000; anti-DNA-PK (PharMingen, clone 4F10C5) 1:500; anti-DNA-PK pT2609 (Rockland) 1:1000; anti-lamin B1 (Biozol, clone ZL-5) 1:1000.

Techniques: Irradiation, SDS Page, Cell Stimulation, Western Blot, Expressing, Derivative Assay

Journal: Molecular Cancer

Article Title: Radiation-induced caveolin-1 associated EGFR internalization is linked with nuclear EGFR transport and activation of DNA-PK

doi: 10.1186/1476-4598-7-69

Figure Lengend Snippet: Stabilization of caveolin1/EGFR complex in cytoplasm after incubation with EGFR inhibitory antibody Erbitux . Confluent A549 cells were treated with human IgG or EGFR inhibitory antibody Erbitux at a concentration of 30 nM for 1 h. Subsequently cells were irradiated with 4 Gy and cytoplasmic and nuclear proteins were isolated at time points given. Proteins were separated by SDS-PAGE and after blotting EGFR protein was quantified with help of a specific antibody. Experiments were performed three times; shown are representative results. Equal protein input for immune-precipitation was documented by showing actin expression within aliquots of input proteins. Expression of specific proteins was quantified by densitometry and shown as fold-induction relative to untreated control.

Article Snippet: The primary antibodies were diluted as follows: anti-EGFR (BD Transduction Laboratories, clone 13) 1:1000; anti EGFR pY845 (nanotools, clone 12A3) 1:1000; anti-EGFR pY992 (abcam, polyclonal) 1:500); anti-EGFR pY1173 (Cell signaling, clone 53A5) 1:1000; anti-phosphotyrosin (Santa Cruz, clone PY20) 1:500); anti-src (Santa Cruz, clone H-12) 1:1000; anti-src Y416 (cell signaling, polyclonal), 1:1000; anti-caveolin-1 (BD Transduction Laboratories, clone 2297) 1:1000; anti-caveolin pY14 (BD Transduction Laboratories, clone 56) 1:1000; anti-DNA-PK (PharMingen, clone 4F10C5) 1:500; anti-DNA-PK pT2609 (Rockland) 1:1000; anti-lamin B1 (Biozol, clone ZL-5) 1:1000.

Techniques: Incubation, Concentration Assay, Irradiation, Isolation, SDS Page, Expressing

Journal: Molecular Cancer

Article Title: Radiation-induced caveolin-1 associated EGFR internalization is linked with nuclear EGFR transport and activation of DNA-PK

doi: 10.1186/1476-4598-7-69

Figure Lengend Snippet: Inhibition of nuclear EGFR transport and phosphorylation of DNA-PK by src inhibitor PP2 . Confluent A549 cells were treated with src inhibitor PP2 at a concentration of 100 nM for 1 h. Subsequently cells were irradiated with 4 Gy and nuclear proteins were isolated at the time points given. Proteins were separated by SDS-PAGE and after blotting protein amounts were quantified by help of specific antibodies. Expression of specific proteins was quantified by densitometry, normalized to lamin B1 and shown as fold-induction relative to untreated control. Experiments were performed three times; shown are representative results.

Article Snippet: The primary antibodies were diluted as follows: anti-EGFR (BD Transduction Laboratories, clone 13) 1:1000; anti EGFR pY845 (nanotools, clone 12A3) 1:1000; anti-EGFR pY992 (abcam, polyclonal) 1:500); anti-EGFR pY1173 (Cell signaling, clone 53A5) 1:1000; anti-phosphotyrosin (Santa Cruz, clone PY20) 1:500); anti-src (Santa Cruz, clone H-12) 1:1000; anti-src Y416 (cell signaling, polyclonal), 1:1000; anti-caveolin-1 (BD Transduction Laboratories, clone 2297) 1:1000; anti-caveolin pY14 (BD Transduction Laboratories, clone 56) 1:1000; anti-DNA-PK (PharMingen, clone 4F10C5) 1:500; anti-DNA-PK pT2609 (Rockland) 1:1000; anti-lamin B1 (Biozol, clone ZL-5) 1:1000.

Techniques: Inhibition, Concentration Assay, Irradiation, Isolation, SDS Page, Expressing

Journal: Diagnostic Pathology

Article Title: Enhancer of zeste homologue 2 (EZH2) is a reliable immunohistochemical marker to differentiate malignant and benign hepatic tumors

doi: 10.1186/1746-1596-7-86

Figure Lengend Snippet: EZH2 staining

Article Snippet: The primary antibody was a mouse monoclonal anti-EZH2 (clone 11/EZH2) from BD Biosciences (San Jose CA, USA) (dilution 1:100).

Techniques: Expressing, Wilms Tumor Assay

Journal: Diagnostic Pathology

Article Title: Enhancer of zeste homologue 2 (EZH2) is a reliable immunohistochemical marker to differentiate malignant and benign hepatic tumors

doi: 10.1186/1746-1596-7-86

Figure Lengend Snippet: EZH2 staining in primary liver tumors. A/HCC, nuclear staining in tumor cells, the surrounding liver is negative; B/hepatocellular adenoma, there is no staining; C/CCC, note the unstained nuclei of the non tumorous bile duct in the center¸D/positively stained highly differentiated CCC (Klatskin tumour). Scale bar for the Figure 1:50 μm.

Article Snippet: The primary antibody was a mouse monoclonal anti-EZH2 (clone 11/EZH2) from BD Biosciences (San Jose CA, USA) (dilution 1:100).

Techniques: Staining

Journal: Scientific Reports

Article Title: Dietary wheat amylase trypsin inhibitors promote features of murine non-alcoholic fatty liver disease

doi: 10.1038/s41598-019-53323-x

Figure Lengend Snippet: ATI feeding increases liver macrophage numbers and their M1- vs M2-type polarization. ( A – C ) Immunohistochemistry and quantitative morphometry for CD68 and YM-1 positive cells (original magnification 40x). ( D ) Ratio of total (CD68+) vs M2-type (Ym-1+) macrophages. ( E ) CD11b+ F4/80+ macrophage subset (% of CD45 positive total immune cells) as determined by FACS analysis. Comparisons by ANOVA; data are means ± SEM for 10 representative sections per mouse and 7–10 mice per group; *p < 0.05, **p < 0.01, ***p < 0.001.

Article Snippet: Tissue was blocked with 5% normal donkey serum, and subsequently incubated with primary antibodies of to CD68 (1:100, Biozol, clone: FA-11), CD86 (1:100, Abcam, cat no: ab119857), and MHC-II (1:100, Abcam, Cat no: 180779) for 2 hrs at room temperature and finally incubated with respective Alexa-flour 488 labelled secondary antibodies.

Techniques: Immunohistochemistry

Journal: Scientific Reports

Article Title: Dietary wheat amylase trypsin inhibitors promote features of murine non-alcoholic fatty liver disease

doi: 10.1038/s41598-019-53323-x

Figure Lengend Snippet: ATI feeding increases hepatic pro-inflammatory and macrophage M1- vs M2-type gene expression. ( A – F ) Hepatic transcript levels of cd68, tnfa, il1b, il6, arg1 and ym1. Comparisons by ANOVA; data are expressed as means ± SEM for 7–10 mice per group; *p < 0.05, **p < 0.01.

Article Snippet: Tissue was blocked with 5% normal donkey serum, and subsequently incubated with primary antibodies of to CD68 (1:100, Biozol, clone: FA-11), CD86 (1:100, Abcam, cat no: ab119857), and MHC-II (1:100, Abcam, Cat no: 180779) for 2 hrs at room temperature and finally incubated with respective Alexa-flour 488 labelled secondary antibodies.

Techniques: Gene Expression

Journal: Scientific Reports

Article Title: Dietary wheat amylase trypsin inhibitors promote features of murine non-alcoholic fatty liver disease

doi: 10.1038/s41598-019-53323-x

Figure Lengend Snippet: Nutritional ATI promote central adipose tissue inflammation. ( A ) Crown like structures (CLS = accumulation of macrophages) in CD68+ stained sections of epididymal adipose tissue in the 4 experimental groups (original magnification 40x), the number of CD68+ CLS as determined by morphometry, and epididymal fat as % of body weight. ( B ) fat weights, and ( C ) transcript levels of cd68, il6 and il1b. Comparisons by ANOVA; data are means ± SEM for 10 representative sections per mouse and 7–10 mice per group; *p < 0.05, **p < 0.01, ***p < 0.001.

Article Snippet: Tissue was blocked with 5% normal donkey serum, and subsequently incubated with primary antibodies of to CD68 (1:100, Biozol, clone: FA-11), CD86 (1:100, Abcam, cat no: ab119857), and MHC-II (1:100, Abcam, Cat no: 180779) for 2 hrs at room temperature and finally incubated with respective Alexa-flour 488 labelled secondary antibodies.

Techniques: Staining

Journal: Scientific Reports

Article Title: Dietary wheat amylase trypsin inhibitors promote features of murine non-alcoholic fatty liver disease

doi: 10.1038/s41598-019-53323-x

Figure Lengend Snippet: ATI feeding increases intestinal macrophage and dendritic cell activation and maturation. ( A–C ) CD68, CD86 and MCH-II expressing cells in the terminal ileum; scale bar: 100 and 50 µm. ( D ) Morphometric quantification of CD68, CD86 and MHC-II positive cells. ( E ) Transcript levels of il1b, tnfα and il6. Comparisons by ANOVA; data are expressed as means ± SEM of 6 mice per group and 5 representative sections per mouse; *p < 0.05, **p < 0.01, ***p < 0.001.

Article Snippet: Tissue was blocked with 5% normal donkey serum, and subsequently incubated with primary antibodies of to CD68 (1:100, Biozol, clone: FA-11), CD86 (1:100, Abcam, cat no: ab119857), and MHC-II (1:100, Abcam, Cat no: 180779) for 2 hrs at room temperature and finally incubated with respective Alexa-flour 488 labelled secondary antibodies.

Techniques: Activation Assay, Expressing