Journal: Acta Neuropathologica

Article Title: Pathogenic implications of cerebrospinal fluid barrier pathology in neuromyelitis optica

doi: 10.1007/s00401-017-1682-1

Figure Lengend Snippet: Patterns of AQP4 and AQP1 expression in the choroid plexus. Normal choroid plexus epithelial cells exhibit variable AQP4 immunoreactivity in the basolateral membrane and cytoplasm ( a ; arrowheads highlight positive cells ), whereas AQP1 is distributed uniformly at the apical membrane in normal choroid plexus epithelial cells ( b ). In hydrocephalus, both AQP4 ( c ) and AQP1 ( d ) immunoreactivity are increased throughout the choroid epithelium. AQP4 immunoreactivity is focally increased in papilloma ( e ; arrowheads ), while AQP1 expression ( f ) is more variable and abnormal. In multiple sclerosis, AQP4 immunoreactivity in the choroid plexus is either normal or increased ( g ; arrowheads ) with more pronounced AQP1 immunoreactivity ( h ). In NMO, AQP4 immunoreactivity is greatly reduced or entirely absent in the choroid plexus ( i ); however, AQP1 immunoreactivity is normal ( j ). Scale bars 20 µm. Immunohistochemistry for AQP4 ( a , c , e , g , i ) and AQP1 ( b , d , f , h , j )

Article Snippet: Immunohistochemistry was performed with the avidin–biotin-complex method as previously reported [ ], using primary antibodies against glial fibrillary acidic protein (GFAP, 1:100, DAKO, Denmark), neurofilament (1:800, steam antigen retrieval with citric acid buffer pH 6.0, DAKO, Denmark), AQP1 (1:250; Santa Cruz), AQP4 (1:250, Sigma-Aldrich, USA), myelin proteolipid protein (PLP, 1:500, Serotec, Oxford, UK), KiM1P (pan-macrophage marker, 1:5000, from Dr. Wolfgang Bruck, Germany), complement C9 neo-antigen (C9neo, monoclonal B7 and polyclonal, 1:200, from Professor Paul Morgan, Cardiff, UK), and human IgG gamma chain (1:200, DAKO, Denmark).

Techniques: Expressing, Membrane, Immunohistochemistry

Journal: PLoS Genetics

Article Title: The Wilms Tumor Gene, Wt1 , Is Critical for Mouse Spermatogenesis via Regulation of Sertoli Cell Polarity and Is Associated with Non-Obstructive Azoospermia in Humans

doi: 10.1371/journal.pgen.1003645

Figure Lengend Snippet: Reproductive tracts of control (A) and Wt1 −/flox ; Cre-ER TM (B) males 3 weeks after Tamoxifen-induced Wt1 ablation. Compared to the control mice, the size of Wt1 −/flox ; Cre- ER TM testes is dramatically reduced (C). (D–I) Cross-sections of control and Wt1 −/flox ; Cre- ER TM testes at 1 (D, G), 2 (E, H), and 3 (F, I) weeks after Tamoxifen treatment. Vacuolization was detected in a small number of tubules at 1 week after Tamoxifen induction (G) and severe epithelial vacuolization (arrows) was noted at 2 weeks after Tamoxifen induction in Wt1 −/flox ; Cre- ER TM testis (H). Massive cell loss with empty tubules (asterisks) was observed in Wt1 −/flox ; Cre- ER TM testis at 3 weeks after Tamoxifen treatment (I).

Article Snippet: IHC analysis of tissues from at least three mice for each genotype was performed using a Vectastain ABC (avidin–biotin–peroxidase) kit (Vector Laboratories, Burlingame, CA) as recommended and using antibodies to WT1 (Santa Cruz, sc-192) and GCNA1 (gift of Dr. George Enders).

Techniques: Control

Journal: PLoS Genetics

Article Title: The Wilms Tumor Gene, Wt1 , Is Critical for Mouse Spermatogenesis via Regulation of Sertoli Cell Polarity and Is Associated with Non-Obstructive Azoospermia in Humans

doi: 10.1371/journal.pgen.1003645

Figure Lengend Snippet: In control testes, the seminiferous tubules are filled with GCNA1 positive germ cells (C), whereas, very few GCNA1 positive germ cells (D, black arrows) are noted in the Wt1 −/flox ; Cre- ER TM testes and some tubules are completely void of germ cells (D, asterisks). Wt1 positive Sertoli cells are present in both control (A) and Wt1 −/flox ; Cre- ER TM (B) testes. The cauda epididymes of control mice are filled with mature sperms (E, black arrows); in contrast, only immature spermatocyte and cell debris (arrow heads) are present in the cauda epididymes of Wt1 −/flox ; Cre-ER TM mice and no mature sperm are detectable (F).

Article Snippet: IHC analysis of tissues from at least three mice for each genotype was performed using a Vectastain ABC (avidin–biotin–peroxidase) kit (Vector Laboratories, Burlingame, CA) as recommended and using antibodies to WT1 (Santa Cruz, sc-192) and GCNA1 (gift of Dr. George Enders).

Techniques: Control

Journal: PLoS Genetics

Article Title: The Wilms Tumor Gene, Wt1 , Is Critical for Mouse Spermatogenesis via Regulation of Sertoli Cell Polarity and Is Associated with Non-Obstructive Azoospermia in Humans

doi: 10.1371/journal.pgen.1003645

Figure Lengend Snippet: Electropherogram showing single-nucleotide mutation of WT1 in patients with non-obstructive azoospermia.

Article Snippet: IHC analysis of tissues from at least three mice for each genotype was performed using a Vectastain ABC (avidin–biotin–peroxidase) kit (Vector Laboratories, Burlingame, CA) as recommended and using antibodies to WT1 (Santa Cruz, sc-192) and GCNA1 (gift of Dr. George Enders).

Techniques: Mutagenesis

Journal: PLoS Genetics

Article Title: The Wilms Tumor Gene, Wt1 , Is Critical for Mouse Spermatogenesis via Regulation of Sertoli Cell Polarity and Is Associated with Non-Obstructive Azoospermia in Humans

doi: 10.1371/journal.pgen.1003645

Figure Lengend Snippet: In control testes, biotin tracer is restricted to the testicular interstitium and the basal compartment of the seminiferous tubules, with no tracer observed in the tubular lumen (A). Biotin tracer is detected along the Sertoli cell plasma membranes from the basement membrane to the lumen in some seminiferous tubules (asterisks) of Wt1 −/flox ; Cre- ER TM testes (B). By TEM a normal BTB structure (E) and apical ES ultrastructure with well-organized actin bundles(C, arrows) is observed in control testes, but Wt1 mutant testes display abnormal apical ES structures (D, arrows) and the BTB structure is disrupted with numerous blisters (F, asterisks and arrow heads).

Article Snippet: IHC analysis of tissues from at least three mice for each genotype was performed using a Vectastain ABC (avidin–biotin–peroxidase) kit (Vector Laboratories, Burlingame, CA) as recommended and using antibodies to WT1 (Santa Cruz, sc-192) and GCNA1 (gift of Dr. George Enders).

Techniques: Control, Clinical Proteomics, Membrane, Mutagenesis

Journal: PLoS Genetics

Article Title: The Wilms Tumor Gene, Wt1 , Is Critical for Mouse Spermatogenesis via Regulation of Sertoli Cell Polarity and Is Associated with Non-Obstructive Azoospermia in Humans

doi: 10.1371/journal.pgen.1003645

Figure Lengend Snippet: Control SCs display a typical epithelial morphology (A and E), while Wt1 -ablated SCs display a mesenchyme-like morphology (C and E). The tight junctions between SCs were assessed by ZO-1 staining. ZO-1 protein is present along the boundary between control SCs (B, white arrows). In contrast, ZO-1 protein is diffused in the cytosol and not detected along the cell boundary in Wt1 -deficient SCs (D). (F) FITC-dextran flux assays indicated increased permeability in Wt1 -deficient SCs compared to control SCs. * p <0.05.

Article Snippet: IHC analysis of tissues from at least three mice for each genotype was performed using a Vectastain ABC (avidin–biotin–peroxidase) kit (Vector Laboratories, Burlingame, CA) as recommended and using antibodies to WT1 (Santa Cruz, sc-192) and GCNA1 (gift of Dr. George Enders).

Techniques: Control, Staining, Permeability

Journal: PLoS Genetics

Article Title: The Wilms Tumor Gene, Wt1 , Is Critical for Mouse Spermatogenesis via Regulation of Sertoli Cell Polarity and Is Associated with Non-Obstructive Azoospermia in Humans

doi: 10.1371/journal.pgen.1003645

Figure Lengend Snippet: (A) The expression of cell polarity related genes ( E-cadherin, Par6b, Cdc42bp5, Sfn ) and Wnt signaling genes ( Wnt4 and Wnt11 ) is significantly decreased in Wt1 -deficient SCs (white bars). (B) E-cadherin and Espin protein expression is significantly reduced in Wt1 -deficient SCs, whereas the expression of occludin , N-cadherin, Zo-1 , and β-catenin is not changed. (C) The expression of E-cadherin, Par6b, Wnt4, Wnt11 , and Cdc42ep5 in Wt1 -deficient SCs was rescued by transfection with Wt1 expressing adenovirus. (D) The expression of Par6b was induced by Wnt4 and Wnt11 expressing adenovirus in Wt1 -deficient SCs. (E) The expression of E-cadherin was not rescued by Wnt4 and Wnt11 expressing adenovirus in Wt1 -deficient SCs. (F) Wnt4 was knocked down by two different siRNAs (siWnt4-2 and siWnt4-3) in SCs and the expression of E-cadherin and Par6b was significantly decreased compared to control cells (NC) treated with a scrambled siRNA. * p <0.05.

Article Snippet: IHC analysis of tissues from at least three mice for each genotype was performed using a Vectastain ABC (avidin–biotin–peroxidase) kit (Vector Laboratories, Burlingame, CA) as recommended and using antibodies to WT1 (Santa Cruz, sc-192) and GCNA1 (gift of Dr. George Enders).

Techniques: Expressing, Transfection, Control

Journal: PLoS Genetics

Article Title: The Wilms Tumor Gene, Wt1 , Is Critical for Mouse Spermatogenesis via Regulation of Sertoli Cell Polarity and Is Associated with Non-Obstructive Azoospermia in Humans

doi: 10.1371/journal.pgen.1003645

Figure Lengend Snippet: (A) The expression of Wnt4, Wnt11 , and E-cadherin in Wt1 -deficient SCs was rescued by wild type Wt1 expressing adenovirus, but not Wt1 R362Q and Wt1 K386R expressing adenovirus. Co-transfection of Wt1 R362Q and Wt1 K386R in Wt1 -deficient SCs did not affect Wt1 induced Wnt4, Wnt11 , and E-cadherin expression. (B) Schematic image of Wnt4 promoter, black oval indicated the predicted Wt1 binding site. (C) A 211 bp band in Wnt4 promoter was amplified after pulled down with Wt1 antibody in Wt1 expressing adenovirus transfected HepG2 cells, but not in Wt1 R362Q and Wt1 K386R expressing adenovirus transfected HepG2 cells. Histone3 antibody and input was used as positive control, and IgG and vector was used as negative control.

Article Snippet: IHC analysis of tissues from at least three mice for each genotype was performed using a Vectastain ABC (avidin–biotin–peroxidase) kit (Vector Laboratories, Burlingame, CA) as recommended and using antibodies to WT1 (Santa Cruz, sc-192) and GCNA1 (gift of Dr. George Enders).

Techniques: Expressing, Cotransfection, Binding Assay, Amplification, Transfection, Positive Control, Plasmid Preparation, Negative Control

Journal: Cancer Science

Article Title: Prognostic significance of pre B cell leukemia transcription factor 2 (PBX2) expression in non‐small cell lung carcinoma

doi: 10.1111/j.1349-7006.2009.01156.x

Figure Lengend Snippet: Effect of pre B cell leukemia transcription factor 1 (PBX1) and PBX2 on valosin‐containing protein (VCP) expression evaluated by siRNA and RT‐PCR analysis on non‐small cell lung cancer (NSCLC) cell lines PC14 and A549. (A) Relative amounts of PBX1 and VCP mRNA to GAPDH mRNA were shown. Decrease in PBX1 expression level by siRNA did not affect the amount of VCP mRNA. (B) Relative amounts of PBX2 and VCP mRNA were shown. The amount of VCP mRNA significantly reduced when PBX2 expression level decreased. *P < 0.05 by Student's test.

Article Snippet: To evaluate the specificity of primary antibodies, the pre‐absorption of anti‐PBX2 and anti‐VCP antibodies with the peptide of PBX2 and VCP (Santa Cruz Biotechnologies; 1:5), respectively, were performed.

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction

Journal: Cancer Science

Article Title: Prognostic significance of pre B cell leukemia transcription factor 2 (PBX2) expression in non‐small cell lung carcinoma

doi: 10.1111/j.1349-7006.2009.01156.x

Figure Lengend Snippet: Analysis of pre B cell leukemia transcription factor 2 (PBX2) binding to the 5′‐flanking region of the human valosin‐containing protein (VCP) gene. ChIP assay was performed using non‐small cell lung cancer (NSCLC) cells. Input DNA sample with (lane A) or without (lane B) the addition of anti‐PBX2 antibody was immunoprecipitated with anti‐PBX2 (lane C) or preimmune IgG serum (lane D). Precipitated DNA fragments were PCR‐amplified with primers specific for the VCP promoter region. PCR products were separated on 2% agarose gel and stained with ethidium bromide. Positive band in the lane of the sample precipitated with anti‐PBX2 antibody (lane A) was seen at the same size with those of input DNA (lanes C and D). M; molecular weight marker.

Article Snippet: To evaluate the specificity of primary antibodies, the pre‐absorption of anti‐PBX2 and anti‐VCP antibodies with the peptide of PBX2 and VCP (Santa Cruz Biotechnologies; 1:5), respectively, were performed.

Techniques: Binding Assay, Immunoprecipitation, Amplification, Agarose Gel Electrophoresis, Staining, Molecular Weight, Marker

Journal: Cancer Science

Article Title: Prognostic significance of pre B cell leukemia transcription factor 2 (PBX2) expression in non‐small cell lung carcinoma

doi: 10.1111/j.1349-7006.2009.01156.x

Figure Lengend Snippet: Pre B cell leukemia transcription factor 2 (PBX2) overexpression up‐regulated luciferase activity of valosin‐containing protein (VCP) promoter in non‐small cell lung cancer (NSCLC) cells of A549 (A) and PC14 (B). The luciferase acivity showed a significant decrease with the transfection of the plasmid mutated or deleted at four PBX2 binding sites. When PBX2 expression plasmid was co‐transfected, the reporter activity significantly increased. Bars represent mean ± SE of at least three independent experiments. *Significant difference (P < 0.05).

Article Snippet: To evaluate the specificity of primary antibodies, the pre‐absorption of anti‐PBX2 and anti‐VCP antibodies with the peptide of PBX2 and VCP (Santa Cruz Biotechnologies; 1:5), respectively, were performed.

Techniques: Over Expression, Luciferase, Activity Assay, Transfection, Plasmid Preparation, Binding Assay, Expressing

Journal: Cancer Science

Article Title: Prognostic significance of pre B cell leukemia transcription factor 2 (PBX2) expression in non‐small cell lung carcinoma

doi: 10.1111/j.1349-7006.2009.01156.x

Figure Lengend Snippet: Comparison of pre B cell leukemia transcription factor 2 (PBX2) protein level determined by Western blot (WB) and immunohistochemistry (IHC). (A) Amount of PBX2 and histone‐H3 proteins in tissues from non‐small cell lung cancer (NSCLC) was determined by WB. (B) The relative amount of PBX2 protein in cases with high and low expression by IHC.

Article Snippet: To evaluate the specificity of primary antibodies, the pre‐absorption of anti‐PBX2 and anti‐VCP antibodies with the peptide of PBX2 and VCP (Santa Cruz Biotechnologies; 1:5), respectively, were performed.

Techniques: Comparison, Western Blot, Immunohistochemistry, Expressing

Journal: Cancer Science

Article Title: Prognostic significance of pre B cell leukemia transcription factor 2 (PBX2) expression in non‐small cell lung carcinoma

doi: 10.1111/j.1349-7006.2009.01156.x

Figure Lengend Snippet: Verification of the specificity of anti‐pre B cell leukemia transcription factor 2 (PBX2) and anti‐valosin‐containing protein (VCP) antibodies. Immunohistochemistry with pre‐absorbed anti‐PBX2 (A), non‐absorbed anti‐PBX2 (B), pre‐absorbed anti‐VCP (C), non‐absorbed anti‐VCP (D) are shown. × 200.

Article Snippet: To evaluate the specificity of primary antibodies, the pre‐absorption of anti‐PBX2 and anti‐VCP antibodies with the peptide of PBX2 and VCP (Santa Cruz Biotechnologies; 1:5), respectively, were performed.

Techniques: Immunohistochemistry

Journal: Cancer Science

Article Title: Prognostic significance of pre B cell leukemia transcription factor 2 (PBX2) expression in non‐small cell lung carcinoma

doi: 10.1111/j.1349-7006.2009.01156.x

Figure Lengend Snippet: Low and high expression of pre B cell leukemia transcription factor 2 (PBX2) by immunohistochemistry. Avidin‐biotin‐peroxidase complex method, ×200.

Article Snippet: To evaluate the specificity of primary antibodies, the pre‐absorption of anti‐PBX2 and anti‐VCP antibodies with the peptide of PBX2 and VCP (Santa Cruz Biotechnologies; 1:5), respectively, were performed.

Techniques: Expressing, Immunohistochemistry, Avidin-Biotin Assay

Journal: Cancer Science

Article Title: Prognostic significance of pre B cell leukemia transcription factor 2 (PBX2) expression in non‐small cell lung carcinoma

doi: 10.1111/j.1349-7006.2009.01156.x

Figure Lengend Snippet: Relationship between PBX2 expression and clinicopathologic factors in 206 patients with non‐small cell lung cancer

Article Snippet: To evaluate the specificity of primary antibodies, the pre‐absorption of anti‐PBX2 and anti‐VCP antibodies with the peptide of PBX2 and VCP (Santa Cruz Biotechnologies; 1:5), respectively, were performed.

Techniques: Expressing, Adjuvant

Journal: Cancer Science

Article Title: Prognostic significance of pre B cell leukemia transcription factor 2 (PBX2) expression in non‐small cell lung carcinoma

doi: 10.1111/j.1349-7006.2009.01156.x

Figure Lengend Snippet: Univariate analysis of clinicopathologic factors for disease‐free and overall survival in 206 patients with non‐small cell lung cancer

Article Snippet: To evaluate the specificity of primary antibodies, the pre‐absorption of anti‐PBX2 and anti‐VCP antibodies with the peptide of PBX2 and VCP (Santa Cruz Biotechnologies; 1:5), respectively, were performed.

Techniques: Expressing, Adjuvant

Journal: Cancer Science

Article Title: Prognostic significance of pre B cell leukemia transcription factor 2 (PBX2) expression in non‐small cell lung carcinoma

doi: 10.1111/j.1349-7006.2009.01156.x

Figure Lengend Snippet: Multivariate analysis of clinicopathologic factors for disease‐free and overall survival in 206 patients with non‐small cell lung cancer

Article Snippet: To evaluate the specificity of primary antibodies, the pre‐absorption of anti‐PBX2 and anti‐VCP antibodies with the peptide of PBX2 and VCP (Santa Cruz Biotechnologies; 1:5), respectively, were performed.

Techniques: Expressing

Journal: Cancer Science

Article Title: Prognostic significance of pre B cell leukemia transcription factor 2 (PBX2) expression in non‐small cell lung carcinoma

doi: 10.1111/j.1349-7006.2009.01156.x

Figure Lengend Snippet: Relationship between PBX2 expression and clinicopathologic factors in 173 non‐small cell lung cancer patients with non‐adjuvant therapy

Article Snippet: To evaluate the specificity of primary antibodies, the pre‐absorption of anti‐PBX2 and anti‐VCP antibodies with the peptide of PBX2 and VCP (Santa Cruz Biotechnologies; 1:5), respectively, were performed.

Techniques: Expressing

Journal: Cancer Science

Article Title: Prognostic significance of pre B cell leukemia transcription factor 2 (PBX2) expression in non‐small cell lung carcinoma

doi: 10.1111/j.1349-7006.2009.01156.x

Figure Lengend Snippet: Disease‐free and overall survival (DFS and OS) rates of patients with high and low expression of pre B cell leukemia transcription factor 2 (PBX2) and valosin‐containing protein (VCP) in non‐small cell lung cancer patients without adjuvant therapy. Significant difference was observed between patients with high and low expression of PBX2 (A and B) and VCP (C and D) for DFS and OS.

Article Snippet: To evaluate the specificity of primary antibodies, the pre‐absorption of anti‐PBX2 and anti‐VCP antibodies with the peptide of PBX2 and VCP (Santa Cruz Biotechnologies; 1:5), respectively, were performed.

Techniques: Expressing, Adjuvant

Journal: Cancer Science

Article Title: Prognostic significance of pre B cell leukemia transcription factor 2 (PBX2) expression in non‐small cell lung carcinoma

doi: 10.1111/j.1349-7006.2009.01156.x

Figure Lengend Snippet: Univariate analysis of clinicopathologic factors for disease‐free and overall survival in 173 non‐small cell lung cnacer patients with non‐adjuvant therapy

Article Snippet: To evaluate the specificity of primary antibodies, the pre‐absorption of anti‐PBX2 and anti‐VCP antibodies with the peptide of PBX2 and VCP (Santa Cruz Biotechnologies; 1:5), respectively, were performed.

Techniques: Expressing

Journal: Cancer Science

Article Title: Prognostic significance of pre B cell leukemia transcription factor 2 (PBX2) expression in non‐small cell lung carcinoma

doi: 10.1111/j.1349-7006.2009.01156.x

Figure Lengend Snippet: Multivariate analysis of clinicopathologic factors for disease‐free and overall survival in 173 non‐small cell lung cancer patients with non‐adjuvant therapy

Article Snippet: To evaluate the specificity of primary antibodies, the pre‐absorption of anti‐PBX2 and anti‐VCP antibodies with the peptide of PBX2 and VCP (Santa Cruz Biotechnologies; 1:5), respectively, were performed.

Techniques: Expressing

Journal: Cancer Science

Article Title: Prognostic significance of pre B cell leukemia transcription factor 2 (PBX2) expression in non‐small cell lung carcinoma

doi: 10.1111/j.1349-7006.2009.01156.x

Figure Lengend Snippet: Disease‐free and overall survival (DFS and OS) rates of patients with high and low expression of pre B cell leukemia transcription factor 2 (PBX2) and VCP at Stage I disease of pathologic tumor‐node‐metastasis (pTNM) classification. Significant difference in survival was not observed between patients with high and low PBX2 expression (A and B), but significant difference was observed between high and low VCP expression for DFS and OS (C and D).

Article Snippet: To evaluate the specificity of primary antibodies, the pre‐absorption of anti‐PBX2 and anti‐VCP antibodies with the peptide of PBX2 and VCP (Santa Cruz Biotechnologies; 1:5), respectively, were performed.

Techniques: Expressing