Journal: Scientific Reports

Article Title: STAT1 and m 6 A-mediated IL15RA upregulation promotes metastasis via ZEB1/NF-κΒ axis in ccRCC

doi: 10.1038/s41598-025-21947-x

Figure Lengend Snippet: IL15RA acts as a promising clinical biomarker for cancers. (A). The correlation between IL15RA expression and IL15, IL2, IL2RA, IL2RG. (B). Paired expression of IL15RA in 33 types of tumors. (C). Univariate Cox analysis of IL15RA and OS, DSS, DFI, PFI at pan-cancer level. (D). Kaplan-Meier analysis of the relationship between IL15RA expression and survival features of tumors in TCGA database, OS Survival Map. (E). OS KMplot analysis of BRCA, KIRP, LAML, LIHC, MESO, PAAD, SKCM, UVM. (F). RFS Survival Map. (G). Diagnosis analysis of IL15RA.ROC analysis of the relationship between IL15RA and clinical diagnosis of tumors in TCGA. IL15RA can better predict cancer diagnosis in KICH, KIRC, CHOL, HNSC, GBM, READ, STAD, and UCEC. (H). Analysis of the expression difference of the IL15RA gene in samples of different clinical stages in each tumor. (I). Analysis of the expression difference of the IL15RA gene in samples of different clinical grades in each tumor. (J). Analysis of the expression difference of the IL15RA gene in samples of different clinical T stages in each tumor. (K). Analysis of the expression difference of the IL15RA gene in samples of different clinical N stages in each tumor. (L). Analysis of the expression difference of the IL15RA gene in samples of different clinical M stages in each tumor. * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001.

Article Snippet: For IL15RA silencing, cells were transfected with IL15RA Human shRNA Plasmid Kit (Locus ID 3601, OriGene, USA).

Techniques: Biomarker Discovery, Expressing

Journal: Scientific Reports

Article Title: STAT1 and m 6 A-mediated IL15RA upregulation promotes metastasis via ZEB1/NF-κΒ axis in ccRCC

doi: 10.1038/s41598-025-21947-x

Figure Lengend Snippet: IL15RA is associated with cancer-related pathways, biological processes and immunity potency. (A). Heatmap showing the Kyoto Encyclopedia of Genes and Genomes pathways related to IL15RA based on GSEA Color ranging from blue to red represents the NES value. (B). Heatmap showing the biological processes related to IL15RA based on GSEA. Color ranging from blue to red represents the NES value. (C). Heatmap showing the correlation between IL15RA expression and 14 kinds of phenotypes based on the CancerSEA database. (D). Immune-related analysis of IL15RA at pan-cancer level. IL15RA expression with immunescore. (E). IL15RA expression with immuinhibitor. (F). IL15RA expression with immustimulator. (G). IL15RA expression with checkpoint. (H). IL15RA expression with chemokine. (I). IL15RA expression with immune-cells. * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001.

Article Snippet: For IL15RA silencing, cells were transfected with IL15RA Human shRNA Plasmid Kit (Locus ID 3601, OriGene, USA).

Techniques: Expressing

Journal: Scientific Reports

Article Title: STAT1 and m 6 A-mediated IL15RA upregulation promotes metastasis via ZEB1/NF-κΒ axis in ccRCC

doi: 10.1038/s41598-025-21947-x

Figure Lengend Snippet: Immunotherapy prediction, mutation and other modification analysis of IL15RA. (A). Immunotherapy response of IL15RA in vivo tumor model in murine ICB therapy cohorts. (B). Immunotherapy response of IL15RA in vitro cell lines. (C). The association between IL15RA and IPS scores using the TCIA database. (D). Biomarker relevance of IL15RA compared to standardized biomarkers on tumor immune evasion in ICB therapy cohorts. (E). Mutation and other modification analysis of IL15RA.Genetic alteration frequency of IL15RA. (F). Correlation between methylation and IL15RA expression. (G). Expression correlation of DNA methyltransferase genes with IL15RA. (H). Correlation of CNV with IL15RA expression. (I). Correlation between mismatch repair genes and IL15RA expression. (J). Correlation analysis of IL15RA and m1A genes. (K). Correlation analysis of IL15RA and m6A genes. (L). Correlation analysis of IL15RA and m5C genes. * P < 0.05; ** P < 0.01; *** P < 0.001.

Article Snippet: For IL15RA silencing, cells were transfected with IL15RA Human shRNA Plasmid Kit (Locus ID 3601, OriGene, USA).

Techniques: Mutagenesis, Modification, In Vivo, In Vitro, Biomarker Discovery, Methylation, Expressing

Journal: Scientific Reports

Article Title: STAT1 and m 6 A-mediated IL15RA upregulation promotes metastasis via ZEB1/NF-κΒ axis in ccRCC

doi: 10.1038/s41598-025-21947-x

Figure Lengend Snippet: Expression, prognosis and single-cell analysis of IL15RA in ccRCC. (A). IL15RA expression between normal and tumor in TCGA and GTEx datasets from GEPIA2. (B). OS analysis of IL15RA in KIRC in UALCAN. (C). OS analysis of IL15RA in KIRP in UALCAN. (D). Expression analysis of IL15RA between normal and tumor from GSE16441 , GSE16449 , GSE36895 , GSE66270 , GSE66271 , GSE66272 , GSE71963 and GSE76351 . (E). Single-cell analysis of IL15RA.Single-cell analysis of IL15RA in kidney cancer. (F-G). Single-cell analysis of IL15RA in ccRCC datasets GSE159115 . (H). Expression analysis of IL15RA in GSE159115 between normal and tumor tissues. (I). Expression analysis of IL15RA in GSE159115 between IL15RA- and IL15RA + cells. (J). GSEA analysis of IL15RA- and IL15RA + cells in GSE159115 based on different cell types. * P < 0.05, ** P < 0.01., *** P < 0.001.

Article Snippet: For IL15RA silencing, cells were transfected with IL15RA Human shRNA Plasmid Kit (Locus ID 3601, OriGene, USA).

Techniques: Expressing, Single-cell Analysis

Journal: Scientific Reports

Article Title: STAT1 and m 6 A-mediated IL15RA upregulation promotes metastasis via ZEB1/NF-κΒ axis in ccRCC

doi: 10.1038/s41598-025-21947-x

Figure Lengend Snippet: The role of IL15RA in genetic and epigenetic regulation, cell migration of ccRCC. (A-B). Western blotting results of IL15RA in cell lines and quantification statistical bar graph. (C). Dual luciferase assay indicated that STAT1 modulates IL15RA transcription activity in ACHN cell line. (D). Overexpression of METTL3 upregulates IL15RA expression in OSRC-2 and ACHN cell lines. (E-F). qRT-PCR results of IL15RA mRNA in OSRC-2 and ACHN cell lines with METTL3 overexpression. (G-H). MeRIP-PCR results of IL15RA mRNA in OSRC-2 and ACHN cell lines with METTL3 overexpression. (I). SRAMP online tool predicted m 6 A binding sites. (J). Dual luciferase assay indicated that site 1219 is the m6A binding site of IL15RA mRNA that modulates IL15RA transcription activity in HEK 293 T cell line. (K). RNA decay assay results of IL15RA mRNA in ACHN cell line with METTL3 overexpression. (L-M). Wound closure test results of IL15RA overexpression and silencing in cell lines and quantification statistical bar graph. (N-O). Transwell migration assay results of IL15RA overexpression and silencing in cell lines and quantification statistical bar graph. * P < 0.05; ** P < 0.01; *** P < 0.001.

Article Snippet: For IL15RA silencing, cells were transfected with IL15RA Human shRNA Plasmid Kit (Locus ID 3601, OriGene, USA).

Techniques: Migration, Western Blot, Luciferase, Activity Assay, Over Expression, Expressing, Quantitative RT-PCR, Binding Assay, Transwell Migration Assay

Journal: Scientific Reports

Article Title: STAT1 and m 6 A-mediated IL15RA upregulation promotes metastasis via ZEB1/NF-κΒ axis in ccRCC

doi: 10.1038/s41598-025-21947-x

Figure Lengend Snippet: IL15RA potentiates ccRCC Metastasis in vivo. (A). OSRC-2 cells with IL15RA overexpression or IL15RA knockdown. (B). Tumor grafts got in vivo. (C). The weight comparison of tumor grafts in B. (D). The tumor volumes measured each week for tumor proliferation. (E). Representative samples for metastasis comparison. (F). Comparison of metastatic foci by naked eye in different groups. * P < 0.05; ** P < 0.01; *** P < 0.001.

Article Snippet: For IL15RA silencing, cells were transfected with IL15RA Human shRNA Plasmid Kit (Locus ID 3601, OriGene, USA).

Techniques: In Vivo, Over Expression, Knockdown, Comparison

Journal: Scientific Reports

Article Title: STAT1 and m 6 A-mediated IL15RA upregulation promotes metastasis via ZEB1/NF-κΒ axis in ccRCC

doi: 10.1038/s41598-025-21947-x

Figure Lengend Snippet: IL15RA potentiates metastasis via NF-κΒ/ZEB1 Axis in ccRCC. (A). Western blotting result of p-p65 protein expression in IL15RA overexpression with or without JSH-23. (B). Quantification analysis of IL15RA protein expression in different groups of A. (C). Quantification analysis of p-p65 protein expression in different groups of A. (D). Western blotting result of p-p65 protein expression in IL15RA knockdown with or without NF-κΒ activator-1. (E). Quantification analysis of IL15RA protein expression in different groups of D. (F). Quantification analysis of p-p65 protein expression in different groups of D. (G). Expression of EMT associated genes in IL15RA overexpression condition in OSRC-2 and ACHN cell lines. (H). Vectors including possible binding sites of p-p65 for ZEB1 promoter region were constructed for dual luciferase assays. (I). Results of luciferase assay performed with ZEB1 full promoter region vector and JSH-23 or NF-κΒ activator-1 in both cell lines. (J). Results of luciferase assay performed with different ZEB1 promoter region vector and IL15RA overexpression in both cell lines. (K). The schematic map for IL15RA regulation in ccRCC. * P < 0.05; ** P < 0.01; *** P < 0.001.

Article Snippet: For IL15RA silencing, cells were transfected with IL15RA Human shRNA Plasmid Kit (Locus ID 3601, OriGene, USA).

Techniques: Western Blot, Expressing, Over Expression, Knockdown, Binding Assay, Construct, Luciferase, Plasmid Preparation

Journal: Scientific Reports

Article Title: STAT1 and m 6 A-mediated IL15RA upregulation promotes metastasis via ZEB1/NF-κΒ axis in ccRCC

doi: 10.1038/s41598-025-21947-x

Figure Lengend Snippet: Drug sensitivity analysis of the low-IL15RAand high-IL15RA groups in TCGA-KIRC. The drug sensitivity showed significant differences between the low-IL15RA and high-IL15RA groups for sunitinib, pyrimethamine, cisplatin, bosutinib, imatinib, cytarabine, paclitaxel gefitinib, lenalidomide.

Article Snippet: For IL15RA silencing, cells were transfected with IL15RA Human shRNA Plasmid Kit (Locus ID 3601, OriGene, USA).

Techniques:

Journal: Molecular Neurodegeneration

Article Title: VCP regulates early tau seed amplification via specific cofactors

doi: 10.1186/s13024-024-00783-z

Figure Lengend Snippet: List of VCP cofactors and their proposed functions

Article Snippet: SYVN1 Human siRNA Oligo Duplex (Locus ID 84447) , OriGene , SR325336.

Techniques: Membrane, Ubiquitin Proteomics

Journal: Molecular Neurodegeneration

Article Title: VCP regulates early tau seed amplification via specific cofactors

doi: 10.1186/s13024-024-00783-z

Figure Lengend Snippet: List of Reagents

Article Snippet: SYVN1 Human siRNA Oligo Duplex (Locus ID 84447) , OriGene , SR325336.

Techniques: Protease Inhibitor, Cell Culture, Transfection, Magnetic Beads, Sequencing, Modification

Journal: Molecular Medicine Reports

Article Title: NLRP3 inflammasome expression affects immune cell infiltration and clinical prognosis in Helicobacter pylori infection‑associated gastric cancer

doi: 10.3892/mmr.2025.13550

Figure Lengend Snippet: NLRP inflammasome expression profile in GC tumors, and NLRP3 expression levels across several tumor and normal tissues. (A) Expression of the NLRP members in gastric cancer data (STAD) of The Cancer Genome Atlas. Expression of the NLRP members in GC datasets (B) GSE36968 and (C) GSE26253 from Gene Expression Omnibus datasets. (D) Differential expression levels of NLRP3 in different tumors based on data obtained from TIMER2.0 datasets. (E) Representative western blotting results of NLRP3 in tissues from patients with GC. (F) Histogram of NLRP3 expression in samples from 10 patients with GC. *P<0.05, **P<0.01, ***P<0.001. GC, gastric cancer; NLRP, nucleotide-binding oligomerization domain, leucine rich repeat and pyrin domain containing.

Article Snippet: C11027-2; Guangzhou RiboBio Co., Ltd.) at 42°C for 30 min. TaqMan Gene Expression Assay primers (cat. no. 4331182; Thermo Fisher Scientific, Inc.) were used for NLRP3 (assay ID Hs00918080_g1) and GAPDH (assay ID Hs02786624_g1) expression.

Techniques: Expressing, Gene Expression, Quantitative Proteomics, Western Blot, Binding Assay

Journal: Molecular Medicine Reports

Article Title: NLRP3 inflammasome expression affects immune cell infiltration and clinical prognosis in Helicobacter pylori infection‑associated gastric cancer

doi: 10.3892/mmr.2025.13550

Figure Lengend Snippet: NLRP3 gene expression in GC tissues with or without Helicobacter pylori infection. (A) Heatmap of genes associated with H. pylori infection in GC tissues from The Cancer Genome Atlas. (B) Volcano plot of differentially expressed genes associated with H. pylori infection in GC tissues. (C) mRNA expression levels of NLRP3 in GC and normal tissues. (D) Comparison of NLRP3 expression in GC tissues from patients with H. pylori (−) and H. pylori (+) GC by IHC. (E) IHC staining of NLRP3 in GC tissues with H. pylori infection. (F) IHC staining of NLRP3 in GC tissues without H. pylori infection. (G) IHC staining of NLRP3 normal gastric mucosa tissues. *P<0.05, ***P<0.001. FC, fold change; FDR, false discovery rate; GC, gastric cancer; IHC, immunohistochemistry; NLRP, nucleotide-binding oligomerization domain, leucine rich repeat and pyrin domain containing.

Article Snippet: C11027-2; Guangzhou RiboBio Co., Ltd.) at 42°C for 30 min. TaqMan Gene Expression Assay primers (cat. no. 4331182; Thermo Fisher Scientific, Inc.) were used for NLRP3 (assay ID Hs00918080_g1) and GAPDH (assay ID Hs02786624_g1) expression.

Techniques: Gene Expression, Infection, Expressing, Comparison, Immunohistochemistry, Binding Assay

Journal: Molecular Medicine Reports

Article Title: NLRP3 inflammasome expression affects immune cell infiltration and clinical prognosis in Helicobacter pylori infection‑associated gastric cancer

doi: 10.3892/mmr.2025.13550

Figure Lengend Snippet: Analysis of hub genes. (A) Protein-protein interaction network of NLRP3. (B) Expression levels of seven related genes in patients with gastric cancer in The Cancer Genome Atlas database. *P<0.05, **P<0.01, ***P<0.001. Survival curve analyses of (C) IL-1B, (D) PYCARD, (E) MAVS, (F) HSP90AA1, (G) TXNIP, (H) NEK7 and (I) BRCC3 genes in gastric cancer. NLRP, nucleotide-binding oligomerization domain, leucine rich repeat and pyrin domain containing; ns, not significant.

Article Snippet: C11027-2; Guangzhou RiboBio Co., Ltd.) at 42°C for 30 min. TaqMan Gene Expression Assay primers (cat. no. 4331182; Thermo Fisher Scientific, Inc.) were used for NLRP3 (assay ID Hs00918080_g1) and GAPDH (assay ID Hs02786624_g1) expression.

Techniques: Expressing, Binding Assay

Journal: Molecular Medicine Reports

Article Title: NLRP3 inflammasome expression affects immune cell infiltration and clinical prognosis in Helicobacter pylori infection‑associated gastric cancer

doi: 10.3892/mmr.2025.13550

Figure Lengend Snippet: Survival curves of patients with high and low NLRP3 expression in GC. (A) A 5-year OS curve of patients with GC. (B) Upregulated expression of NLRP3 in the Kaplan-Meier Plotter database was associated with a worse 10-year OS. (C) Progression-free survival and (D) post-progression survival analysis based on NLRP3 expression using the Kaplan-Meier Plotter database. GC, gastric cancer; NLRP, nucleotide-binding oligomerization domain, leucine rich repeat and pyrin domain containing; OS, overall survival.

Article Snippet: C11027-2; Guangzhou RiboBio Co., Ltd.) at 42°C for 30 min. TaqMan Gene Expression Assay primers (cat. no. 4331182; Thermo Fisher Scientific, Inc.) were used for NLRP3 (assay ID Hs00918080_g1) and GAPDH (assay ID Hs02786624_g1) expression.

Techniques: Expressing, Binding Assay

Journal: Molecular Medicine Reports

Article Title: NLRP3 inflammasome expression affects immune cell infiltration and clinical prognosis in Helicobacter pylori infection‑associated gastric cancer

doi: 10.3892/mmr.2025.13550

Figure Lengend Snippet: Results of CIBERSORT analysis on the infiltration of different immune cells affected by NLRP3 expression in GC tissues. (A) Proportion of immune infiltration in GC tumor tissues vs. normal tissues. (B) Box plot of the proportions of immune cells in GC tissues. (C) Heatmap of the correlation of different immune cell infiltrates in GC tissues. Blue indicates a positive correlation, and red indicates a negative correlation. (D) Types of immune cell infiltrates primarily affected by NLRP3 and other genes. Red indicates a positive correlation and indicates a negative correlation. *P<0.05, **P<0.01, ***P<0.001. GC, gastric cancer; NLRP, nucleotide-binding oligomerization domain, leucine rich repeat and pyrin domain containing.

Article Snippet: C11027-2; Guangzhou RiboBio Co., Ltd.) at 42°C for 30 min. TaqMan Gene Expression Assay primers (cat. no. 4331182; Thermo Fisher Scientific, Inc.) were used for NLRP3 (assay ID Hs00918080_g1) and GAPDH (assay ID Hs02786624_g1) expression.

Techniques: Expressing, Binding Assay

Journal: Molecular Medicine Reports

Article Title: NLRP3 inflammasome expression affects immune cell infiltration and clinical prognosis in Helicobacter pylori infection‑associated gastric cancer

doi: 10.3892/mmr.2025.13550

Figure Lengend Snippet: GSEA of NLRP3-influenced gene signaling function pathways in gastric cancer tissues. (A) Top 15 signaling pathways significantly affected by NLRP3. (B) Signaling pathways activated and suppressed by NLRP3. (C) GSEA of signaling pathways activated (E2F targets gene set) or inhibited (oxidative phosphorylation gene set) by NLRP3. (D) NLRP3 GSEA mountain range maps. (E) NLRP3 GSEA bubble plots. GSEA, Gene Set Enrichment Analysis; NLRP, nucleotide-binding oligomerization domain, leucine rich repeat and pyrin domain containing.

Article Snippet: C11027-2; Guangzhou RiboBio Co., Ltd.) at 42°C for 30 min. TaqMan Gene Expression Assay primers (cat. no. 4331182; Thermo Fisher Scientific, Inc.) were used for NLRP3 (assay ID Hs00918080_g1) and GAPDH (assay ID Hs02786624_g1) expression.

Techniques: Protein-Protein interactions, Phospho-proteomics, Binding Assay

Journal: Molecular Medicine Reports

Article Title: NLRP3 inflammasome expression affects immune cell infiltration and clinical prognosis in Helicobacter pylori infection‑associated gastric cancer

doi: 10.3892/mmr.2025.13550

Figure Lengend Snippet: Correlation of NLRP3 with immune checkpoints. (A) Scatterplots showing the correlation between immune checkpoint genes and NLRP3 expression. (B) Single gene co-expression heat map of NLRP3 with immune checkpoint genes. ***P<0.001. NLRP, nucleotide-binding oligomerization domain, leucine rich repeat and pyrin domain containing.

Article Snippet: C11027-2; Guangzhou RiboBio Co., Ltd.) at 42°C for 30 min. TaqMan Gene Expression Assay primers (cat. no. 4331182; Thermo Fisher Scientific, Inc.) were used for NLRP3 (assay ID Hs00918080_g1) and GAPDH (assay ID Hs02786624_g1) expression.

Techniques: Expressing, Binding Assay

Journal: Molecular Medicine Reports

Article Title: NLRP3 inflammasome expression affects immune cell infiltration and clinical prognosis in Helicobacter pylori infection‑associated gastric cancer

doi: 10.3892/mmr.2025.13550

Figure Lengend Snippet: NLRP3 expression levels and macrophage polarization in gastric cancer. Scatter plots of (A) tumor-associated macrophage, (B) monocyte, (C) M1 macrophage and (D) M2 macrophage infiltration levels based on NLRP3 expression. NLRP, nucleotide-binding oligomerization domain, leucine rich repeat and pyrin domain containing.

Article Snippet: C11027-2; Guangzhou RiboBio Co., Ltd.) at 42°C for 30 min. TaqMan Gene Expression Assay primers (cat. no. 4331182; Thermo Fisher Scientific, Inc.) were used for NLRP3 (assay ID Hs00918080_g1) and GAPDH (assay ID Hs02786624_g1) expression.

Techniques: Expressing, Binding Assay

Journal: Molecular Medicine Reports

Article Title: NLRP3 inflammasome expression affects immune cell infiltration and clinical prognosis in Helicobacter pylori infection‑associated gastric cancer

doi: 10.3892/mmr.2025.13550

Figure Lengend Snippet: High NLRP3 expression promotes immune cell infiltration. (A) Representative IHC staining images of immune cell surface markers. (B) Enlarged IHC image of the M2 macrophage marker CD206. (C) Histogram of immunohistochemical cell count scores. *P<0.05, ***P<0.001. NLRP, nucleotide-binding oligomerization domain, leucine rich repeat and pyrin domain containing; IHC, immunohistochemistry.

Article Snippet: C11027-2; Guangzhou RiboBio Co., Ltd.) at 42°C for 30 min. TaqMan Gene Expression Assay primers (cat. no. 4331182; Thermo Fisher Scientific, Inc.) were used for NLRP3 (assay ID Hs00918080_g1) and GAPDH (assay ID Hs02786624_g1) expression.

Techniques: Expressing, Immunohistochemistry, Marker, Immunohistochemical staining, Cell Counting, Binding Assay

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Induction of TRAIL- and TNF-?-dependent Apoptosis in Human Monocyte-derived Dendritic Cells by Microfilariae of Brugia Malayi 1

doi:

Figure Lengend Snippet: Pre-exposure to mf upregulates mRNA expression of TRAIL, TL1A, and TNF-α and induces production of soluble TL1A and TNF-α but downregulates mRNA expression of DR3 in DC. A, Results are expressed as 1/ΔCT for DC unexposed or mf-exposed (DC/mf). The higher the 1/average ΔCT, the higher the expression of the gene. Each line represents an independent donor. B, Data are expressed as ng/ml for DC unexposed or exposed to mf for 48 h (DC/mf). Each line represents an independent donor.

Article Snippet: The assay ID numbers from Applied Biosystems for the genes tested in this study is listed below: Gene name Assay ID TRAIL Hs00234356_m1 TNF-α Hs00174128_m1 TL1A Hs00270802_s1 DR3 Hs00600930_g1 BID Hs00609632_m1 BCL2 Hs00153350_m1 FADD Hs00938709_m1 TRADD Hs00182558_m1 TRAF2 Hs00184186_m1 FLIP Hs00153439_ml Caspase 8 Hs00236278_m1 Open in a separate window Real-time RT-PCR Relative transcript levels were determined by the formula: 1/(Cttarget - Ctcontrol), where CT is the threshold cycle during the exponential phase of amplification.

Techniques: Expressing

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Induction of TRAIL- and TNF-?-dependent Apoptosis in Human Monocyte-derived Dendritic Cells by Microfilariae of Brugia Malayi 1

doi:

Figure Lengend Snippet: DC are sensitive to TRAIL- (but not TL1A)–induced cell death. DC were cultured either alone or with A recombinant TRAIL in the presence of anti-TRAIL antibody or isotype control antibody for 48 h or B recombinant TL1A in the absence or presence of cyclohexamide. Cells were harvested, and viability of cells was measured by PI+ dead cells using flow cytometry.

Article Snippet: The assay ID numbers from Applied Biosystems for the genes tested in this study is listed below: Gene name Assay ID TRAIL Hs00234356_m1 TNF-α Hs00174128_m1 TL1A Hs00270802_s1 DR3 Hs00600930_g1 BID Hs00609632_m1 BCL2 Hs00153350_m1 FADD Hs00938709_m1 TRADD Hs00182558_m1 TRAF2 Hs00184186_m1 FLIP Hs00153439_ml Caspase 8 Hs00236278_m1 Open in a separate window Real-time RT-PCR Relative transcript levels were determined by the formula: 1/(Cttarget - Ctcontrol), where CT is the threshold cycle during the exponential phase of amplification.

Techniques: Cell Culture, Recombinant, Control, Flow Cytometry

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Induction of TRAIL- and TNF-?-dependent Apoptosis in Human Monocyte-derived Dendritic Cells by Microfilariae of Brugia Malayi 1

doi:

Figure Lengend Snippet: Pre-exposure to mf results in upregulation of Bid mRNA expression but downregulation of cytoplasmic tBid protein. mf upregulate cytochrome c protein expression in DC in a TRAIL- and TNF-α-dependent manner and induce cleavage of 35-kDa and 17-kDa active caspase 9. A, Results are expressed as 1/average ΔCT in unexposed and mf-exposed DC (DC/mf) as individual lines representing each of 6 to 14 independent donors. The higher the 1/average ΔCT, the higher the expression of the gene. B, Total Bid (22 kDa) and tBid (15 kDa) in DC unexposed and exposed to mf for 48 h using western blot analysis in one representative of 12 different donors. Graph demonstrates the tBid:tubulin ratio for each of 12 donors. C, Cytochrome c (14 kDa) protein expression in DC unexposed and exposed to mf for 48 h in 1 representative of 11 different donors and in the absence or presence of anti-TRAIL-R2, anti-TRAIL-R1, or anti-TNF-α using immunoblot analysis. Graphs demonstrate the cytochrome c to tubulin ratio for each of 11 donors or of 4 donors. D, 47-kDa pro-caspase 9 was cleaved to 35-kDa and 17-kDa active fragments of DC exposed to mf for 48 h in one of three independent donors. For positive control in all immunoblots, caspase 3 cell extracts were used (see Materials and Method section).

Article Snippet: The assay ID numbers from Applied Biosystems for the genes tested in this study is listed below: Gene name Assay ID TRAIL Hs00234356_m1 TNF-α Hs00174128_m1 TL1A Hs00270802_s1 DR3 Hs00600930_g1 BID Hs00609632_m1 BCL2 Hs00153350_m1 FADD Hs00938709_m1 TRADD Hs00182558_m1 TRAF2 Hs00184186_m1 FLIP Hs00153439_ml Caspase 8 Hs00236278_m1 Open in a separate window Real-time RT-PCR Relative transcript levels were determined by the formula: 1/(Cttarget - Ctcontrol), where CT is the threshold cycle during the exponential phase of amplification.

Techniques: Expressing, Western Blot, Positive Control