Journal: Cancer Research

Article Title: Insulin-like Growth Factor–Binding Protein-2 Is a Target for the Immunomodulation of Breast Cancer

doi: 10.1158/0008-5472.can-07-5891

Figure Lengend Snippet: Figure 5. IGFBP-2 protein expression in tumor cells derived from neu transgenic mice. IGFBP-2 monoclonal antibody staining of MMC cells. A, secondary antibody alone. B, IGFBP-2 protein expression. Magnification, 400. Black bar, 20 mm. C, top, Western blot of 10 Ag SKBR3, MCF-7, T98G, MMC, and neu transgenic mouse spleen lysate. Arrow, 40-kDa level of molecular weight (MW) marker (36-kDa IGFBP-2). Bottom, h-actin expression in 10 Ag of the same lysates.

Article Snippet: Commercially provided T98G cell lysate (Santa Cruz Biotechnology) was used as a positive control.

Techniques: Expressing, Derivative Assay, Transgenic Assay, Staining, Western Blot, Molecular Weight, Marker

Journal: American Journal of Cancer Research

Article Title: Claudin 4 enhances the malignancy of glioma cells via NNAT/Wnt signaling

doi:

Figure Lengend Snippet: CLND4 promotes malignancy in glioma cells. A. Expression levels (mRNA) of CLDN4 in normal tissues (n=5) and glioma tissues (n=156); data derived from TCGA database. B. Representative images of immunohistochemical staining for CLDN4 in para-carcinomatous tissues (n=15), early-stage glioma tissues (stage I-II, n=15), and late-stage glioma tissues (stage III-IV, n=15); scale bar =50 μm. C. Kaplan-Meier survival curve for high and low expression of CLDN4 in glioma patients (n=513); data from TCGA database. D. Western blotting for CLDN4 in LN229/T98G cells transfected with empty vector or a CLDN4-overexpression vector. E. Cell proliferation assay (CCK-8 assay) for LN229/T98G cells transfected with empty vector or CLDN4-overexpression vector. T98G, P<0.05, 72 hours; LN229, P<0.01, 72 hours. F. Cell migration assay (Transwell assay) for LN229/T98G cells transfected with empty vector or CLDN4-overexpression vector; scale bar =100 μm. G. Colony formation assay for LN229/T98G cells transfected with empty vector or CLDN4-overexpression vector.

Article Snippet: Transfected LN229 and T98G cells overexpressing CLDN4 were obtained from GenScript ProBio (China); the same cells transfected with the empty vector (without the CLDN4 gene) were also obtained from the same source.

Techniques: Expressing, Derivative Assay, Immunohistochemical staining, Staining, Western Blot, Transfection, Plasmid Preparation, Over Expression, Proliferation Assay, CCK-8 Assay, Cell Migration Assay, Transwell Assay, Colony Assay

Journal: American Journal of Cancer Research

Article Title: Claudin 4 enhances the malignancy of glioma cells via NNAT/Wnt signaling

doi:

Figure Lengend Snippet: CLDN4-stimulated NNAT upregulation. A. The top 30 upregulated genes in patients with gliomas that had high CLDN4 expression levels as compared to those with gliomas having low CLDN4 expression levels; data derived from TCGA database (n=513). B. Western blotting for CLDN4 and NNAT in LN229/T98G cells transfected with empty vector or CLDN4-overexpression vector. C. Cell proliferation assays for LN229/T98G cells transfected with empty vector or CLDN4-overexpression vector treated with scramble or NNAT siRNAs. LN229, vector vs vector + siRNA, P>0.05, OE vs OE + siRNA, P<0.05, 72 hours; T98G, vector vs vector + siRNA, P>0.05, OE vs OE + siRNA, P<0.01, 72 hours. D. Cell migration assay for LN229/T98G cells transfected with empty vector or CLDN4-overexpression vector treated with scramble or NNAT siRNAs. E. Colony formation assays for LN229/T98G cells transfected with empty vector or CLDN4-overexpression vector treated with scramble or NNAT siRNAs. F. Representative images of immunohistochemical staining for CLND4 in early-stage glioma tissues (stage I-II, n=15) and late-stage glioma tissues (stage III-IV, n=15); scale bar =100 μm.

Article Snippet: Transfected LN229 and T98G cells overexpressing CLDN4 were obtained from GenScript ProBio (China); the same cells transfected with the empty vector (without the CLDN4 gene) were also obtained from the same source.

Techniques: Expressing, Derivative Assay, Western Blot, Transfection, Plasmid Preparation, Over Expression, Cell Migration Assay, Immunohistochemical staining, Staining

Journal: American Journal of Cancer Research

Article Title: Claudin 4 enhances the malignancy of glioma cells via NNAT/Wnt signaling

doi:

Figure Lengend Snippet: The CLDN4/NNAT axis modulates glioma progression through Wnt signaling. (A and B) The results of the KEGG enrichment analysis for the major signaling pathways involved in glioma progression in 513 glioma patients divided into CLDN4-high/low (A) or NNAT-high/low (B) groups. (C) Western blotting for Wnt1, Wnt2, and Wnt3A in LN229 cells transfected with empty vector or CLDN4-overexpression vector. (D) Western blotting for Wnt3A in LN229/T98G cells transfected with empty vector or CLDN4-overexpression vector treated with scramble or NNAT siRNAs. (E) Cell proliferation assays for LN229/T98G cells transfected with empty vector or CLDN4-overexpression vector treated with PBS or pyrvinium pamoate (10 nM). LN229, vector vs vector + PP, P>0.05, OE vs OE + PP, P<0.05, 72 hours; T98G, vector vs vector + PP, P>0.05, OE vs OE + PP, P<0.05, 72 hours. (F) Cell migration assays for LN229/T98G cells transfected with empty vector or CLDN4-overexpression vector treated with PBS or pyrvinium pamoate (10 nM). (G) Colony formation assays for LN229/T98G cells transfected with empty vector or CLDN4-overexpression vector treated with PBS or pyrvinium pamoate (10 nM).

Article Snippet: Transfected LN229 and T98G cells overexpressing CLDN4 were obtained from GenScript ProBio (China); the same cells transfected with the empty vector (without the CLDN4 gene) were also obtained from the same source.

Techniques: Protein-Protein interactions, Western Blot, Transfection, Plasmid Preparation, Over Expression, Migration

Journal: American Journal of Cancer Research

Article Title: Claudin 4 enhances the malignancy of glioma cells via NNAT/Wnt signaling

doi:

Figure Lengend Snippet: CLDN4 facilitates glioma growth in vivo. A. Western blotting for CLDN4 in primary glioma cells derived from 6 patients. B. Representative images of PDOs derived from CLDN4-high and -low glioma tissues; scale bar =50 μm. C. Western blotting for Wnt3A and NNAT in PDOs derived from CLDN4-high and -low glioma tissues. D. Representative images for immunofluorescence staining for Ki67 in PDOs derived from CLDN4-high and -low glioma tissues; scale bar =50 μm. E. Tumor volumes of subcutaneous tumors formed by LN229 cells transfected with empty vector or CLDN4-overexpression vector (n=5 in each group). F. Kaplan-Meier survival curve for mice with tumors formed by LN229 cells transfected with empty vector or CLDN4-overexpression vector (n=5 in each group).

Article Snippet: Transfected LN229 and T98G cells overexpressing CLDN4 were obtained from GenScript ProBio (China); the same cells transfected with the empty vector (without the CLDN4 gene) were also obtained from the same source.

Techniques: In Vivo, Western Blot, Derivative Assay, Immunofluorescence, Staining, Transfection, Plasmid Preparation, Over Expression

Journal: Cell Death & Disease

Article Title: Decreased expression of the Augmenter of Liver Regeneration results in increased apoptosis and oxidative damage in human-derived glioma cells

doi: 10.1038/cddis.2012.25

Figure Lengend Snippet: Effect of rALR treatment on H 2 O 2 -induced ROS levels on glioma cells. Human T98G glioma cells, treated with H 2 O 2 , were incubated with different concentrations of rALR (1–100 ng/ml). The means±S.D. of six triplicate independent experiments are shown

Article Snippet: Human-derived T98G glioma cells (Interlab Cell Line Collection, Genoa, Italy) were cultured in Dulbecco's modified Eagle's medium (DMEM) (Sigma-Aldrich) supplemented with 10% (v/v) inactivated FBS (PAA Laboratories GmbH, Pasching, Austria), 2 mM L -glutamine (Sigma-Aldrich), 100 μ g/ml penicillin and 100 μ g/ml streptomycin (Sigma-Aldrich,) at 37°C in 5% CO 2 .

Techniques: Incubation

Journal: Cell Death & Disease

Article Title: Decreased expression of the Augmenter of Liver Regeneration results in increased apoptosis and oxidative damage in human-derived glioma cells

doi: 10.1038/cddis.2012.25

Figure Lengend Snippet: Effect of siRNA/ALR treatment on ALR expression. ALR expression in siRNA/ALR-treated and control human T98G glioma cells, determined using western blot analysis ( a ) and confocal microscopy ( b , green colour), are reported. The data shown are means±S.D. from five independent experiments

Article Snippet: Human-derived T98G glioma cells (Interlab Cell Line Collection, Genoa, Italy) were cultured in Dulbecco's modified Eagle's medium (DMEM) (Sigma-Aldrich) supplemented with 10% (v/v) inactivated FBS (PAA Laboratories GmbH, Pasching, Austria), 2 mM L -glutamine (Sigma-Aldrich), 100 μ g/ml penicillin and 100 μ g/ml streptomycin (Sigma-Aldrich,) at 37°C in 5% CO 2 .

Techniques: Expressing, Western Blot, Confocal Microscopy

Journal: Cell Death & Disease

Article Title: Decreased expression of the Augmenter of Liver Regeneration results in increased apoptosis and oxidative damage in human-derived glioma cells

doi: 10.1038/cddis.2012.25

Figure Lengend Snippet: Effect of siRNA/ALR treatment on clusterin expression. Clusterin expression in siRNA/ALR-treated and control human T98G glioma cells, determined using western blot analysis ( a , upper right) and confocal microscopy ( b , green colour), are reported. The data shown are means±S.D. from three independent experiments

Article Snippet: Human-derived T98G glioma cells (Interlab Cell Line Collection, Genoa, Italy) were cultured in Dulbecco's modified Eagle's medium (DMEM) (Sigma-Aldrich) supplemented with 10% (v/v) inactivated FBS (PAA Laboratories GmbH, Pasching, Austria), 2 mM L -glutamine (Sigma-Aldrich), 100 μ g/ml penicillin and 100 μ g/ml streptomycin (Sigma-Aldrich,) at 37°C in 5% CO 2 .

Techniques: Expressing, Western Blot, Confocal Microscopy

Journal: Cell Death & Disease

Article Title: Decreased expression of the Augmenter of Liver Regeneration results in increased apoptosis and oxidative damage in human-derived glioma cells

doi: 10.1038/cddis.2012.25

Figure Lengend Snippet: Effect of siRNA/ALR treatment on apoptotic gene expression. bcl-2 and bax ( a ), caspase-9 ( b ) and activated caspase-3 ( c ) expression in siRNA/ALR-treated and control human T98G glioma cells, determined using western blot analysis, are reported. The data shown are means±S.D. from five independent experiments

Article Snippet: Human-derived T98G glioma cells (Interlab Cell Line Collection, Genoa, Italy) were cultured in Dulbecco's modified Eagle's medium (DMEM) (Sigma-Aldrich) supplemented with 10% (v/v) inactivated FBS (PAA Laboratories GmbH, Pasching, Austria), 2 mM L -glutamine (Sigma-Aldrich), 100 μ g/ml penicillin and 100 μ g/ml streptomycin (Sigma-Aldrich,) at 37°C in 5% CO 2 .

Techniques: Expressing, Western Blot