Journal: Viruses

Article Title: Pediatric and Adult High-Grade Glioma Stem Cell Culture Models Are Permissive to Lytic Infection with Parvovirus H-1

doi: 10.3390/v8050138

Figure Lengend Snippet: Stem cell marker expression and 50% inhibition (LD50) oncolytic parvovirus H-1 (H-1PV) virus doses in high-grade glioma (HGG) neurospheres.

Article Snippet: Adult GBM neurospheres (NCH 421k, NCH421R, NCH644) and pediatric GBM secondary neurospheres (SF-188NS and KNS-42NS) were cultured in Dulbecco's Modified Eagle Medium (DMEM) containing 20% BIT serum-free supplement, basic fibroblast growth factor (bFGF, 20 ng/mL), and epidermal growth factor (EGF, 20 ng/mL), (all from Provitro GmbH, Berlin, Germany), as previously described [ ].

Techniques: Marker, Expressing, Inhibition, Virus

Journal: Viruses

Article Title: Pediatric and Adult High-Grade Glioma Stem Cell Culture Models Are Permissive to Lytic Infection with Parvovirus H-1

doi: 10.3390/v8050138

Figure Lengend Snippet: H-1 parvovirus (H-1PV) initiates replication in adult and pediatric HGG neurospheres. Indicated HGG neurosphere cultures were infected with H-1PV (one PFU per cell) six days post seeding. ( A ) Three days post infection, the initiation of virus replication was measured by fluorescence microscopy after cell immuno-staining for the nonstructural viral protein 1 (NS1) ( green ), co-staining of neurospheres with Hoechst 33342 ( blue ) is shown in merge panels; ( B ) H-1PV virus production was assayed in a time course experiment by quantification of infectious particles as described in .

Article Snippet: Adult GBM neurospheres (NCH 421k, NCH421R, NCH644) and pediatric GBM secondary neurospheres (SF-188NS and KNS-42NS) were cultured in Dulbecco's Modified Eagle Medium (DMEM) containing 20% BIT serum-free supplement, basic fibroblast growth factor (bFGF, 20 ng/mL), and epidermal growth factor (EGF, 20 ng/mL), (all from Provitro GmbH, Berlin, Germany), as previously described [ ].

Techniques: Infection, Virus, Fluorescence, Microscopy, Immunostaining, Staining

Journal: Viruses

Article Title: Pediatric and Adult High-Grade Glioma Stem Cell Culture Models Are Permissive to Lytic Infection with Parvovirus H-1

doi: 10.3390/v8050138

Figure Lengend Snippet: H-1PV infection induces lytic infection of HGG neurospheres in vitro . Lactate dehydrogenase (LDH)-release assays were performed on NCH421k and NCH421R cells at days 3 and 6 after infection with H-1PV at increasing MOIs. Specific lysis is given in relation to completely lysed cells by detergent treatment. Means from six replicates and respective standard errors are displayed.

Article Snippet: Adult GBM neurospheres (NCH 421k, NCH421R, NCH644) and pediatric GBM secondary neurospheres (SF-188NS and KNS-42NS) were cultured in Dulbecco's Modified Eagle Medium (DMEM) containing 20% BIT serum-free supplement, basic fibroblast growth factor (bFGF, 20 ng/mL), and epidermal growth factor (EGF, 20 ng/mL), (all from Provitro GmbH, Berlin, Germany), as previously described [ ].

Techniques: Infection, In Vitro, Lysis

Journal: Viruses

Article Title: Pediatric and Adult High-Grade Glioma Stem Cell Culture Models Are Permissive to Lytic Infection with Parvovirus H-1

doi: 10.3390/v8050138

Figure Lengend Snippet: H-1PV infection suppresses tumorigenicity of HGG neurospheres in vivo. ( A ) NCH421R and ( B ) NCH421k neurosphere cultures were infected with H-1PV (MOI = one or ten PFU/cell) 4 h prior to subcutaneous implantation into the right flank of NOD/SCID mice (10 6 cells per animal). Mock-treated NCH421k cells served as non-treatment controls. Mock-infected NCH421k and NCH421R xenografts were confirmed to preserve a HGG histomorphology (hematoxylin/eosin, 400× magnification). Significant differences in xenograft tumor proliferation between mock infected cells and H-1PV infected cells were only observed in NCH421k xenograft bearing animals ( p < 0.001).

Article Snippet: Adult GBM neurospheres (NCH 421k, NCH421R, NCH644) and pediatric GBM secondary neurospheres (SF-188NS and KNS-42NS) were cultured in Dulbecco's Modified Eagle Medium (DMEM) containing 20% BIT serum-free supplement, basic fibroblast growth factor (bFGF, 20 ng/mL), and epidermal growth factor (EGF, 20 ng/mL), (all from Provitro GmbH, Berlin, Germany), as previously described [ ].

Techniques: Infection, In Vivo

Journal: International Journal of Molecular Sciences

Article Title: Estrogen α and β Receptor Expression in the Various Regions of Resected Glioblastoma Multiforme Tumors and in an In Vitro Model

doi: 10.3390/ijms25074130

Figure Lengend Snippet: Expression of ERα gene ( A ) and ERα protein ( B ) in U87 cells cultured under different conditions. Data are representative of each group cultured in control, nutrient-deficient, hypoxic, and necrotic conditions. Statistical analysis was performed using t -test *, p < 0.005.

Article Snippet: Cell culture of the U87 line (obtained from the European Collection of Authenticated Cell Cultures (ECACC)) was performed under standard conditions of 37 °C, 95% humidity, and 5% CO 2 , according to the manufacturer’s instructions.

Techniques: Expressing, Cell Culture, Control

Journal: International Journal of Molecular Sciences

Article Title: Estrogen α and β Receptor Expression in the Various Regions of Resected Glioblastoma Multiforme Tumors and in an In Vitro Model

doi: 10.3390/ijms25074130

Figure Lengend Snippet: Representative images taken with the FV1000 confocal microscope system (Olympus, Hamburg, Germany) show ERα protein expression in U87 cells cultured under specific conditions: control ( A ), nutrient deficiency ( B ), hypoxia ( C ), and necrotic conditions ( D ). FITC (AR) and DAPI (nuclear) markers were used. Microphotographs were taken at ×20 magnification ( A , B , D ) and ×40 magnification ( C ); scale bar 30 µm.

Article Snippet: Cell culture of the U87 line (obtained from the European Collection of Authenticated Cell Cultures (ECACC)) was performed under standard conditions of 37 °C, 95% humidity, and 5% CO 2 , according to the manufacturer’s instructions.

Techniques: Microscopy, Expressing, Cell Culture, Control

Journal: International Journal of Molecular Sciences

Article Title: Estrogen α and β Receptor Expression in the Various Regions of Resected Glioblastoma Multiforme Tumors and in an In Vitro Model

doi: 10.3390/ijms25074130

Figure Lengend Snippet: Expression of the ERβ gene ( A ) and ERβ protein ( B ) in U87 cells cultured under different conditions. Data are representative of each group cultured in control, nutrient-deficient, hypoxic, and necrotic conditions. Statistical analysis was performed using a t -test * p < 0.05.

Article Snippet: Cell culture of the U87 line (obtained from the European Collection of Authenticated Cell Cultures (ECACC)) was performed under standard conditions of 37 °C, 95% humidity, and 5% CO 2 , according to the manufacturer’s instructions.

Techniques: Expressing, Cell Culture, Control

Journal: International Journal of Molecular Sciences

Article Title: Estrogen α and β Receptor Expression in the Various Regions of Resected Glioblastoma Multiforme Tumors and in an In Vitro Model

doi: 10.3390/ijms25074130

Figure Lengend Snippet: Representative images taken with the FV1000 confocal microscope system (Olympus, Hamburg, Germany) show ERβ protein expression in U87 cells cultured under specific conditions: control ( A ), nutrient deficiency ( B ), hypoxia ( C ), and necrotic conditions ( D ). FITC (AR) and DAPI (nuclear) markers were used. Microphotographs were taken at ×20 magnification ( A , B , D ) and ×40 magnification ( C ); scale bar 30 µm.

Article Snippet: Cell culture of the U87 line (obtained from the European Collection of Authenticated Cell Cultures (ECACC)) was performed under standard conditions of 37 °C, 95% humidity, and 5% CO 2 , according to the manufacturer’s instructions.

Techniques: Microscopy, Expressing, Cell Culture, Control

Journal: OncoTargets and therapy

Article Title: Secretory Pathway Kinase FAM20C , a Marker for Glioma Invasion and Malignancy, Predicts Poor Prognosis of Glioma

doi: 10.2147/OTT.S275452

Figure Lengend Snippet: The development of secretory pathway kinase or kinase-like proteins (SPKKPs) gene signature stratifies the IDH wild type (wt) GBM as two groups with distinct survival. ( A ) The coefficient profiles of 13 SPKKPs genes with the gradual increase of lambda by LASSO regression (TCGA GBM RNA-seq, IDH wt, n = 142). ( B ) LASSO regression analysis with cross-validation method identified a SPKKPs gene signature including 3 members in this family ( FAM20A, FAM20A , and C3orf58 ) with prognostic value in IDH wt GBM (TCGA, n = 142). ( C ) Heatmap showing the association of 13 SPKKPs gene expression with clinicopathologic features in low- and high-risk GBM groups defined by the secretory pathway kinase related gene signature (TCGA GBM RNA-seq: low risk: n = 71, high risk: n = 71, Chi-square test). ( D ) Kaplan–Meier curves describing the survival of IDH wt GBM in low- and high-risk groups defined by secretory pathway-related gene signature (TCGA GBM RNA-seq: low risk: n = 71, high risk: n = 71, Log rank test, P = 0.0312). ( E ) The expression of SPKKPs member genes in different WHO grades of glioma (TCGA RNA-seq: grade II: n = 260, grade III: n = 267, GBM: n = 168, one-way ANOVA). ( F ) The expression of SPKKPs member genes in GBM with different IDH status (TCGA GBM RNA-seq: IDH mutant (mut): n = 11, IDH wt: n = 144, t -test). ( G ) The expression of SPKKPs member genes in all low-grade gliomas (LGG) and IDH -mut LGG with different 1p/19q codeletion (codel) status (TCGA RNA-seq: LGG 1p19q codel: n = 160, LGG 1p19q non-codel: n = 317; LGG with IDH mut 1p19q codel: n = 160, LGG with IDH mut 1p19q non-codel: n = 230, t -test). (ns P > 0.05, * P < 0.05, *** P < 0.001, and **** P < 0.0001).

Article Snippet: Figure 4 FAM20C knockdown suppresses the migration, invasion, and colony formation of GBM cells. ( A ) GSEA with TCGA GBM RNA-seq dataset disclosed a significant enrichment of cell adhesion- and immune response-related phenotypes in GBM patients with high FAM20C expression. ( B ) Heat maps describing the association between FAM20C expression and cell adhesion and negative immune-regulation‐related genes in TCGA GBM RNA-seq dataset (Chi-square test). ( C ) KEGG analysis was performed on genes with a correlation coefficient greater than 0.3 (Spearman analysis) with FAM20C in TCGA GBM RNA-seq dataset. ( D ) qPCR analyses of FAM20C, MMP2, and MMP9 mRNA expression in LN229 cells transfected with siRNA targeting FAM20C or a non-targeting control (n = 4, t -test). ( E ) Transwell assays demonstrating FAM20C knock-down inhibited the migration (upper panel) and invasion (lower panel) capabilities of LN229 cells (n = 10, t -test). ( ) The colony formation assay showing FAM20C knock-down significantly inhibited the colony formation capability of LN229 cells. (n = 6, t -test). (* P < 0.05, *** P < 0.001, and **** P < 0.0001).

Techniques: RNA Sequencing, Biomarker Discovery, Gene Expression, Expressing, Mutagenesis

Journal: OncoTargets and therapy

Article Title: Secretory Pathway Kinase FAM20C , a Marker for Glioma Invasion and Malignancy, Predicts Poor Prognosis of Glioma

doi: 10.2147/OTT.S275452

Figure Lengend Snippet: Integrative transcriptomic analyses identify FAM20C as the core member of secretory pathway kinase or kinase-like proteins (SPKKPs) family in glioma. ( A ) The protein interaction analysis among SPKKPs member genes with STRING ( https://string-db.org ). Dark green and pink lines represent known interactions. Green, red, and blue lines represent predicted interactions. Light green, black and gray lines represent other interactions. ( B and C ) Spearman correlation ( B ), the circle size represents correlation strength, and the color represents the positive (orange) or negative (blue) correlation and the univariate Cox regression analyses ( C ) of secretory pathway-related genes in TCGA glioma RNA-seq dataset. ( D ) Oncomine analysis of FAM20C expression in indicated cancers (The number in red represents the number of datasets demonstrating elevated FAM20C expression in indicated cancers. The number in blue represents the number of datasets showing decreased FAM20C expression in indicated cancers. The intensity of the color means the level of P value).

Article Snippet: Figure 4 FAM20C knockdown suppresses the migration, invasion, and colony formation of GBM cells. ( A ) GSEA with TCGA GBM RNA-seq dataset disclosed a significant enrichment of cell adhesion- and immune response-related phenotypes in GBM patients with high FAM20C expression. ( B ) Heat maps describing the association between FAM20C expression and cell adhesion and negative immune-regulation‐related genes in TCGA GBM RNA-seq dataset (Chi-square test). ( C ) KEGG analysis was performed on genes with a correlation coefficient greater than 0.3 (Spearman analysis) with FAM20C in TCGA GBM RNA-seq dataset. ( D ) qPCR analyses of FAM20C, MMP2, and MMP9 mRNA expression in LN229 cells transfected with siRNA targeting FAM20C or a non-targeting control (n = 4, t -test). ( E ) Transwell assays demonstrating FAM20C knock-down inhibited the migration (upper panel) and invasion (lower panel) capabilities of LN229 cells (n = 10, t -test). ( ) The colony formation assay showing FAM20C knock-down significantly inhibited the colony formation capability of LN229 cells. (n = 6, t -test). (* P < 0.05, *** P < 0.001, and **** P < 0.0001).

Techniques: RNA Sequencing, Expressing

Journal: OncoTargets and therapy

Article Title: Secretory Pathway Kinase FAM20C , a Marker for Glioma Invasion and Malignancy, Predicts Poor Prognosis of Glioma

doi: 10.2147/OTT.S275452

Figure Lengend Snippet: FAM20C is associated with progressive malignancy and unfavorable prognosis in glioma. ( A ) Representative immunohistochemical images of FAM20C staining in clinical glioma samples (Scale bar, 50 μm; grade II: n = 3, grade III: n = 7, grade IV: n=28). ( B ) Kaplan–Meier curve evaluating the correlation between FAM20C protein expression and GBM patients’ survival (FAM20C low vs high, low n = 11, high n = 17, P = 0.0241; Log rank test). ( C ) The analyses of FAM20C expression in non-tumor and different grade glioma samples (TCGA glioma RNA-seq: non-tumor, n = 5; grade II: n = 130; grade III: n = 133; GBM: n = 80, one-way ANOVA). ( D ) Kaplan–Meier curves of FAM20C expression and the survival of different grade glioma in TCGA. (left panel: grade II, low n = 130, high n = 130, P = 0.710; middle panel: grade III, low n = 133, high n = 134, P = 0.0069; right panel: GBM, low n = 80, high n = 80, P = 0.0013, Log rank test). ( E ) The analysis of FAM20C expression in non-tumor and GBM samples using data from Clinical Proteomic Tumor Analysis Consortium (CPTAC, non-tumor, n = 10; GBM, n = 100, P < 0.0001, t -test). ( F ) Kaplan–Meier curves of FAM20C expression and GBM patients’ survival in CPTAC. (high: n = 46, low: n = 47, P = 0.0032, Log rank test) ( G ) The analysis of FAM20C expression in different regions of GBM with data from the IVY GBM Altas Project ( http://glioblastoma.alleninstitute.org/ ). ( H ) Representative immunohistochemical images of FAM20C staining in peri-necrotic region of GBM. (Scale bar, 50 μm). ( I ) The analyses of FAM20C expression in different subtypes GBM (TCGA GBM RNA-seq: classical n = 48; mesenchymal n = 65; proneural n = 18, one-way ANOVA). ( J ) The receiver operator characteristic (ROC) curve describing the sensitivity and specificity of FAM20C as a marker for mesenchymal (n = 65) vs other subtypes (classical n = 48, and proneural n = 18) in TCGA. ( K ) FAM20C expression analysis in GBM with different IDH status (TCGA GBM RNA-seq: IDH mut, n = 11; IDH wt, n = 143, P < 0.0001, t -test). ( L ) Kaplan‐Meier curve describing the association between FAM20C expression and GBM IDH wt patients’ 2-year survival (TCGA, low n = 71, high n = 71, P = 0.0288, Log rank test). ( M ) The ROC curves comparing the sensitivity and specificity of FAM20C as a prognostic marker for glioma patients in TCGA (left panel: 3‐year; right: 5‐year). ( N and O ) Kaplan–Meier curves describing the association between FAM20C expression and TCGA GBM patients’ survival with or without radiation ( N ) or chemotherapy ( O ) (N: low without radiation n = 39, low with radiation n = 43; high without radiation n = 32, high with radiation n = 50; ( O ) low without chemotherapy n = 22, low with chemotherapy n = 61; high without chemotherapy n = 18, high with chemotherapy n = 66, Log rank test). (ns P > 0.05, * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001).

Article Snippet: Figure 4 FAM20C knockdown suppresses the migration, invasion, and colony formation of GBM cells. ( A ) GSEA with TCGA GBM RNA-seq dataset disclosed a significant enrichment of cell adhesion- and immune response-related phenotypes in GBM patients with high FAM20C expression. ( B ) Heat maps describing the association between FAM20C expression and cell adhesion and negative immune-regulation‐related genes in TCGA GBM RNA-seq dataset (Chi-square test). ( C ) KEGG analysis was performed on genes with a correlation coefficient greater than 0.3 (Spearman analysis) with FAM20C in TCGA GBM RNA-seq dataset. ( D ) qPCR analyses of FAM20C, MMP2, and MMP9 mRNA expression in LN229 cells transfected with siRNA targeting FAM20C or a non-targeting control (n = 4, t -test). ( E ) Transwell assays demonstrating FAM20C knock-down inhibited the migration (upper panel) and invasion (lower panel) capabilities of LN229 cells (n = 10, t -test). ( ) The colony formation assay showing FAM20C knock-down significantly inhibited the colony formation capability of LN229 cells. (n = 6, t -test). (* P < 0.05, *** P < 0.001, and **** P < 0.0001).

Techniques: Immunohistochemical staining, Staining, Expressing, RNA Sequencing, Marker

Journal: OncoTargets and therapy

Article Title: Secretory Pathway Kinase FAM20C , a Marker for Glioma Invasion and Malignancy, Predicts Poor Prognosis of Glioma

doi: 10.2147/OTT.S275452

Figure Lengend Snippet: FAM20C knockdown suppresses the migration, invasion, and colony formation of GBM cells. ( A ) GSEA with TCGA GBM RNA-seq dataset disclosed a significant enrichment of cell adhesion- and immune response-related phenotypes in GBM patients with high FAM20C expression. ( B ) Heat maps describing the association between FAM20C expression and cell adhesion and negative immune-regulation‐related genes in TCGA GBM RNA-seq dataset (Chi-square test). ( C ) KEGG analysis was performed on genes with a correlation coefficient greater than 0.3 (Spearman analysis) with FAM20C in TCGA GBM RNA-seq dataset. ( D ) qPCR analyses of FAM20C, MMP2, and MMP9 mRNA expression in LN229 cells transfected with siRNA targeting FAM20C or a non-targeting control (n = 4, t -test). ( E ) Transwell assays demonstrating FAM20C knock-down inhibited the migration (upper panel) and invasion (lower panel) capabilities of LN229 cells (n = 10, t -test). ( F ) The colony formation assay showing FAM20C knock-down significantly inhibited the colony formation capability of LN229 cells. (n = 6, t -test). (* P < 0.05, *** P < 0.001, and **** P < 0.0001).

Article Snippet: Figure 4 FAM20C knockdown suppresses the migration, invasion, and colony formation of GBM cells. ( A ) GSEA with TCGA GBM RNA-seq dataset disclosed a significant enrichment of cell adhesion- and immune response-related phenotypes in GBM patients with high FAM20C expression. ( B ) Heat maps describing the association between FAM20C expression and cell adhesion and negative immune-regulation‐related genes in TCGA GBM RNA-seq dataset (Chi-square test). ( C ) KEGG analysis was performed on genes with a correlation coefficient greater than 0.3 (Spearman analysis) with FAM20C in TCGA GBM RNA-seq dataset. ( D ) qPCR analyses of FAM20C, MMP2, and MMP9 mRNA expression in LN229 cells transfected with siRNA targeting FAM20C or a non-targeting control (n = 4, t -test). ( E ) Transwell assays demonstrating FAM20C knock-down inhibited the migration (upper panel) and invasion (lower panel) capabilities of LN229 cells (n = 10, t -test). ( ) The colony formation assay showing FAM20C knock-down significantly inhibited the colony formation capability of LN229 cells. (n = 6, t -test). (* P < 0.05, *** P < 0.001, and **** P < 0.0001).

Techniques: Knockdown, Migration, RNA Sequencing, Expressing, Transfection, Control, Colony Assay

Journal: OncoTargets and therapy

Article Title: Secretory Pathway Kinase FAM20C , a Marker for Glioma Invasion and Malignancy, Predicts Poor Prognosis of Glioma

doi: 10.2147/OTT.S275452

Figure Lengend Snippet: The screening of FAM20C substrates in GBM distinguishes FN1 as the key substrate interacting with it. ( A ) The analyses of protein interactions between FAM20C and its substrates by STRING webtool ( https://string-db.org ). ( B ) The univariate Cox regression analyses of FAM20C substrates in TCGA GBM RNA-seq dataset. ( C ) Pearson correlation analysis between FAM20C and its substrates (TCGA GBM RNA-seq dataset: circle size represents the correlation strength, and color represents the positive (orange) or negative (blue) correlation). ( D ) Pearson correlation analysis between FAM20C and FN1 in TCGA GBM RNA-seq dataset. ( E ) Three-dimensional binding pattern of FAM20C and FN1 protein obtained by molecular docking simulation. ( F ) The interaction site between FAM20C and FN1.

Article Snippet: Figure 4 FAM20C knockdown suppresses the migration, invasion, and colony formation of GBM cells. ( A ) GSEA with TCGA GBM RNA-seq dataset disclosed a significant enrichment of cell adhesion- and immune response-related phenotypes in GBM patients with high FAM20C expression. ( B ) Heat maps describing the association between FAM20C expression and cell adhesion and negative immune-regulation‐related genes in TCGA GBM RNA-seq dataset (Chi-square test). ( C ) KEGG analysis was performed on genes with a correlation coefficient greater than 0.3 (Spearman analysis) with FAM20C in TCGA GBM RNA-seq dataset. ( D ) qPCR analyses of FAM20C, MMP2, and MMP9 mRNA expression in LN229 cells transfected with siRNA targeting FAM20C or a non-targeting control (n = 4, t -test). ( E ) Transwell assays demonstrating FAM20C knock-down inhibited the migration (upper panel) and invasion (lower panel) capabilities of LN229 cells (n = 10, t -test). ( ) The colony formation assay showing FAM20C knock-down significantly inhibited the colony formation capability of LN229 cells. (n = 6, t -test). (* P < 0.05, *** P < 0.001, and **** P < 0.0001).

Techniques: RNA Sequencing, Binding Assay

Journal: OncoTargets and therapy

Article Title: Secretory Pathway Kinase FAM20C , a Marker for Glioma Invasion and Malignancy, Predicts Poor Prognosis of Glioma

doi: 10.2147/OTT.S275452

Figure Lengend Snippet: FAM20C is associated with the regulation of immune response in GBM. ( A ) Heat maps describing the expression of FAM20C negatively correlated with tumor purity, and positively correlated with immune score and stromal score (TCGA GBM RNA-seq, n = 168, Pearson correlation analysis). ( B ) The analyses of xCell (upper panel) and EPIC scores (lower panel) indicating FAM20C expression pattern in indicated cell populations (TCGA GBM RNA-seq: FAM20C low n = 84; high n = 84, t -test). ( C and D ) t-SNE map color-coded for transcript counts ( C ) and the corresponding single-cell bar plots ( D ) of FAM20C enriched in different cell subpopulations of GBM (SCP393 dataset, https://portals.broadinstitute.org/single_cell/study/SCP393/single-cell-rna-seq-of-adult-and-pediatric-glioblastoma ). ( E ) Transwell assay showing 20 μg/mL FAM20C recombinant protein significantly enhances the migration of THP1 cells. (n = 15, t -test). (ns P > 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001).

Article Snippet: Figure 4 FAM20C knockdown suppresses the migration, invasion, and colony formation of GBM cells. ( A ) GSEA with TCGA GBM RNA-seq dataset disclosed a significant enrichment of cell adhesion- and immune response-related phenotypes in GBM patients with high FAM20C expression. ( B ) Heat maps describing the association between FAM20C expression and cell adhesion and negative immune-regulation‐related genes in TCGA GBM RNA-seq dataset (Chi-square test). ( C ) KEGG analysis was performed on genes with a correlation coefficient greater than 0.3 (Spearman analysis) with FAM20C in TCGA GBM RNA-seq dataset. ( D ) qPCR analyses of FAM20C, MMP2, and MMP9 mRNA expression in LN229 cells transfected with siRNA targeting FAM20C or a non-targeting control (n = 4, t -test). ( E ) Transwell assays demonstrating FAM20C knock-down inhibited the migration (upper panel) and invasion (lower panel) capabilities of LN229 cells (n = 10, t -test). ( ) The colony formation assay showing FAM20C knock-down significantly inhibited the colony formation capability of LN229 cells. (n = 6, t -test). (* P < 0.05, *** P < 0.001, and **** P < 0.0001).

Techniques: Expressing, RNA Sequencing, Transwell Assay, Recombinant, Migration

Journal: Cancer Informatics

Article Title: multiClust : An R-package for Identifying Biologically Relevant Clusters in Cancer Transcriptome Profiles

doi: 10.4137/CIN.S38000

Figure Lengend Snippet: Summary of gene expression datasets used in this study.

Article Snippet: The TCGA Glioblastoma dataset was downloaded from the TCGA Data Portal and was composed of median expression data from three different expression centers: The Broad Institute, University of North Carolina, and the Lawrence Berkley National Laboratory.

Techniques: Gene Expression