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: PLoS Biology

Article Title: Reporters to mark and eliminate basal or luminal epithelial cells in culture and in vivo

doi: 10.1371/journal.pbio.2004049

Figure Lengend Snippet: (A) Cartoon of the K14 promoter–driving expression of tRFP-P2A-TK (K14.tRPT) reporter. (B) Cartoon of the K8 promoter–driving expression of tGFP-P2A-DTR (K8.tGPD) reporter construct. (C) Phase contrast images of K14+, K14−, and K8+ cells grown in monolayer (upper panels, scale bar 100 μm) and 3D (lower panels, scale bar 200 μm) on M/Col-I culture for 4 days. (D) Quantification of invasive structures in C, represented as means ± SEM from 3 independent experiments; * p < 0.001 by unpaired t test. (E) K14+ and K8+ cells were treated with DT (2.5 ng/ml), GCV (1 μg/ml), or media. Column bars indicate the percentage of reporter-positive and negative cells after treatments for the indicated cell lines, as determined by flow cytometry. (F) Immunoblots of lysates from K14+ and K14− reporter cell lines were analyzed for changes in expression of E-cadherin, β-catenin, fibronectin, vimentin, and α-SMA. Blots were also probed with antibodies for GAPDH and α-tubulin as loading controls. Right panel shows quantification of vimentin for 3 independent western blot p = 0.0317 by paired t test; mean ± SD is shown. (G) Immunofluorescence of K14+ (upper panel) and K14− (lower panel) monolayers for vimentin, β-catenin, or detection of endogenous tRFP signal. Scale bar 25 μm. α-smooth muscle actin; DT, diphtheria toxin; DTR, diphtheria toxin receptor; GAPDH, glyceraldehyde 3-phosphate dehydrogenase; GCV, ganciclovir; K, cytokeratin; K8.tGPD, keratin-8 promoter followed by turbo green fluorescent protein and diphtheria toxin receptor; K14.tRPT, keratin-14 promoter followed by a turbo red fluorescent protein and herpes simplex virus thymidine kinase; M/Col-I; 1:1 mixture of Matrigel/Collagen-I; pA, polyadenylation signal sequence; TK, thymidine kinase; tGFP, turbo green fluorescent protein; tRFP, turbo red fluorescent protein

Article Snippet: Primary antibodies to E-cadherin (clone 36) and β-catenin (clone 14) secondary antibody PE-Cy7 streptavidin and conjugated antibody CD24-APC (Clone M1/69) were purchased from BD Transduction Laboratories.

Techniques: Expressing, Construct, Flow Cytometry, Western Blot, Immunofluorescence, Sequencing