af 400 sp tnf α neutralizing antibody r d system  (R&D Systems)

Journal: Cell Death Discovery

Article Title: Pan-caspase inhibitors induce secretion of HIV-1 latency reversal agent lymphotoxin-alpha from cytokine-primed NK cells

doi: 10.1038/s41420-025-02330-1

Figure Lengend Snippet: A TZM-bl cells were treated with different concentrations of TNF or LTα overnight. Activation of the HIV-1 LTR promoter was assessed by subjecting these cells to a luciferase assay. Data points are plotted as mean ± SD from technical triplicates. Two-way ANOVA was performed with comparisons for each cytokine to the unstimulated control. B , C Optimal TNF and LTα concentrations were pre-incubated with various concentrations of the corresponding neutralizing Ab (nAb) against either TNF or LTα. To determine the neutralizing capacity of the nAb, these mixtures were added to TZM-bl cells overnight followed by a luciferase assay (gray bars). TZM-bl cells incubated with only nAb was used as a control (black bars). Data points are plotted as mean ± SD from technical triplicates. Ordinary one-way ANOVA was performed on the depicted comparisons. D , E KHYG-1 cells were incubated with DMSO (0.5%) or Z-VAD-FMK (50 µM) overnight. Supernatants were collected and pre-incubated with concentration ranges of either TNF or LTα nAb before adding to TZM-bl cells overnight followed by a luciferase assay. Data points are plotted as mean ± SD from three individual experiments each with technical triplicates. Two-way ANOVA was performed with comparisons to 0 ng/mL nAb for each treatment. Only comparisons for Z-VAD-FMK treatment are shown. All comparisons for DMSO treatment were non-significant. F KHYG-1 cells were incubated with DMSO (0.5%) or Z-VAD-FMK (50 µM) overnight. Supernatants were collected and pre-incubated with no antibody, LTα nAb (6.5 µg/mL), or IgG2A isotype control (6.5 µg/mL) before adding to TZM-bl cells overnight followed by a luciferase assay. Data points are plotted as mean ± SD from three individual experiments each with technical triplicates. Two-way ANOVA was performed. Only comparisons for Z-VAD-FMK treatment are shown. All comparisons for DMSO treatment were non-significant. (ns, non-significant: ** p < 0.01, **** p < 0.0001).

Article Snippet: Human LTα neutralizing mouse monoclonal antibody (clone 5807R, cat. no. MAB621R), mouse IgG2a isotype control (clone 133304, cat. no. MAB0031), and human NKp30 agonistic mouse monoclonal antibody (clone 210847, cat. no. MAB18491) were purchased from R&D Systems (Minneapolis, MN, USA).

Techniques: Activation Assay, Luciferase, Control, Incubation, Concentration Assay

Journal: JCI insight

Article Title: Cytomegalovirus infection lengthens the cell cycle of granule cell precursors during postnatal cerebellar development.

doi: 10.1172/jci.insight.175525

Figure Lengend Snippet: Figure 9. Treatment with TNF-NAb normalizes cell cycle abnormalities in MCMV-infected mice. (A) Schematic representation of IdU-BrdU dual-labeling protocol in MCMV-infected mice and control mice treated with vehicle (Veh), isotype control antibody (Iso), or TNF-α neutralizing antibody (TNF-NAb). Fixed brain sections were stained for IdU, BrdU, Ki67, and DAPI to measure cell cycle parameters as described in previous figures. (B and C) Brain sections from mice treated with BrdU for 30 minutes were analyzed to measure cell proliferation by quantifying BrdU+ (B) and Ki67+ (C) GCPs in the EGL. (D) Mice were labeled with IdU for 6 hours, followed by BrdU injection 30 minutes prior to harvest- ing brains at P8 to estimate the length of S phase of GCPs in the EGL. (E and F) Mice were pulse labeled with IdU for 22 hours and injected with BrdU 30 minutes prior to harvesting brains at P8 to estimate cell cycle exit (E) and total cell cycle length (F). Data shown as mean ± SD, n = 4–6 mice/experimental group for immunofluorescence. Each data point corresponds to cerebellar EGL from an individual mouse. P values were calculated by using 1-way ANOVA with Tukey’s multiple-comparison test. *P < 0.05; **P < 0.01; ***P < 0.001.

Article Snippet: TNF-α neutralizing antibody (TNF-NAb) treatment Pups were treated daily (P3–P7) by i.p. injection with InVivoMAb rat IgG1 Isotype control anti-trinitrophenol (anti-TNP) (In VivoMAb rat IgG1 Isotype control; anti-TNP; BioXcell; BE0290; clone TNP6A7) or InVivoMAb anti–mouse TNF-α neutralizing antibody (tat IgG1, clone XT3.11, BioXCell) at 500 μg/ mouse/day diluted in PBS.

Techniques: Infection, Labeling, Control, Staining, Injection, Immunofluorescence, Comparison