pa5 19810  (Cell Signaling Technology Inc)

Journal: Journal of Inflammation Research

Article Title: P2X7 Receptor and Heart Function in a Mouse Model of Systemic Inflammation Due to High Fat Diet

doi: 10.2147/JIR.S356038

Figure Lengend Snippet: Heart gene expression profile of key regulators of oxidative stress modulation, endothelial function and chemotactic processes observed in wild-type mice treated with normal (WT-NFD) or high (WT-HFD) fat diet, or P2X7 receptor knockout mice treated with normal (KO-HFD) or high fat diet (KO-HFD). Tumor necrosis factor α (TNFα, A ), Nitric oxide synthase 2, inducible (NOS-2, B ), Monocyte chemotactic protein-1 (MCP-1, C ), Nitric oxide synthase 3, endothelial (NOS-3, D ), Adhesion G protein-coupled receptor E1 (ADGRE1, E ), Intercellular adhesion molecule 1 (ICAM-1, F ) are shown. Relative gene expression was calculated by the 2 −ΔΔCt method and normalized by the β-actin housekeeping gene. Results are reported as arbitrary units (A.U.). Data are presented as mean±SE for at least six animals in each group. Two-way ANOVA with genotype and diet as sources of variation, followed by Tukey’s post-hoc test, was used for multiple comparisons. Statistical significance was set at p < 0.05. ● WT-NFD, ■ WT-HFD, ▲ KO-NFD, ◆ KO-HFD.

Article Snippet: After blocking with BSA 5% in T-tris buffered saline (TBS and Tween-20 0.1%) for 1h at RT, blots were incubated overnight at 4°C with one of the following primary antibodies diluted 1:1000: anti-IL-1β (number 12242), anti-IL-6 (number 12912), and anti-p38 mitogen-activated protein kinase (p38MAPK) (number 9212), anti-p-p38MAPK (number 9211), anti-transforming growth factor-β (TGFβ) (number 3711), anti-monocyte chemoattractant protein-1 (MCP-1) (number 2029) from Cell Signaling Technology (Danvers, MA, USA); anti-NLRP3 (AG-20B-0014) from Adipogen (San Diego, CA, USA); anti-nitric oxide synthase-2 (NOS-2) (sc-651) from Santa Cruz Biotechnology (Dallas, TX, USA), and anti-tumor necrosis factor-α (TNFα) (PA5-19810) from ThermoFisher Scientific.

Techniques: Expressing, Knock-Out

Journal: Journal of Inflammation Research

Article Title: P2X7 Receptor and Heart Function in a Mouse Model of Systemic Inflammation Due to High Fat Diet

doi: 10.2147/JIR.S356038

Figure Lengend Snippet: Heart protein expression of key regulators of oxidative stress modulation, endothelial function and chemotactic processes whose genes were differentially expressed in wild-type mice treated with normal (WT-NFD) or high (WT-HFD) fat diet, or P2X7 receptor knockout mice treated with normal (KO-NFD) or high fat diet (KO-HFD). Tumor necrosis factor α (TNFα, A ), nitric oxide synthase 2, inducible (NOS-2, B ), Monocyte chemotactic protein-1 (MCP-1, C ) are shown. Band intensities were normalized based on reference protein GAPDH and results are expressed as arbitrary units (A.U.). Arrows indicate different MCP-1 isoforms. Data are presented as mean±SE for at least six animals in each group. Two-way ANOVA with genotype and diet as sources of variation, followed by Tukey’s post-hoc test, was used for multiple comparisons. Statistical significance was set at p < 0.05. ● WT-NFD, ■ WT-HFD, ▲ KO-NFD, ◆ KO-HFD.

Article Snippet: After blocking with BSA 5% in T-tris buffered saline (TBS and Tween-20 0.1%) for 1h at RT, blots were incubated overnight at 4°C with one of the following primary antibodies diluted 1:1000: anti-IL-1β (number 12242), anti-IL-6 (number 12912), and anti-p38 mitogen-activated protein kinase (p38MAPK) (number 9212), anti-p-p38MAPK (number 9211), anti-transforming growth factor-β (TGFβ) (number 3711), anti-monocyte chemoattractant protein-1 (MCP-1) (number 2029) from Cell Signaling Technology (Danvers, MA, USA); anti-NLRP3 (AG-20B-0014) from Adipogen (San Diego, CA, USA); anti-nitric oxide synthase-2 (NOS-2) (sc-651) from Santa Cruz Biotechnology (Dallas, TX, USA), and anti-tumor necrosis factor-α (TNFα) (PA5-19810) from ThermoFisher Scientific.

Techniques: Expressing, Knock-Out

Journal: Frontiers in Cellular Neuroscience

Article Title: Successful Treatment of Noise-Induced Hearing Loss by Mesenchymal Stromal Cells: An RNAseq Analysis of Protective/Repair Pathways

doi: 10.3389/fncel.2021.656930

Figure Lengend Snippet: Graphical representation of IPA analysis of up- and down-regulated genes and processes. Using IPA core analysis of down-regulated genes and processes (blue) predicted up-regulation of genes and processes (orange). There is an overall down-regulation of pathways related to synaptogenesis, development of neurons and excitatory signaling. TNFα is a key factor that influences multiple processes and genes.

Article Snippet: For TNFα immunohistochemistry the tissue was incubated for 48 h at 4°C in a humid chamber in TNFα polyclonal antibody (1:500, Thermo Fisher, PA5-19810).

Techniques:

Journal: Frontiers in Cellular Neuroscience

Article Title: Successful Treatment of Noise-Induced Hearing Loss by Mesenchymal Stromal Cells: An RNAseq Analysis of Protective/Repair Pathways

doi: 10.3389/fncel.2021.656930

Figure Lengend Snippet: Expression of TNFα protein (Red A–C ) and semi quantitative RT PCR of TNFα mRNA (D) . Control cochlea showed minimal TNF expression in the hair cells and lateral wall (A) . After sound trauma there was a clear increase in TFN alpha immunolabeling in hair cells, the spiral ganglion (arrow) and the lateral wall (B) . Treatment with MSCs after sound trauma reduced TNFα expression in the lateral wall and outer hair cells but expression remained high in the inner hair cell (arrow C ). Semi quantitative RT-PCR showed an up-regulation of TNF gene expression after sound trauma. This was slightly reduced after MSC treatment but not to control levels (D) .

Article Snippet: For TNFα immunohistochemistry the tissue was incubated for 48 h at 4°C in a humid chamber in TNFα polyclonal antibody (1:500, Thermo Fisher, PA5-19810).

Techniques: Expressing, Quantitative RT-PCR, Control, Immunolabeling, Gene Expression