Journal: Microbiology and Immunology

Article Title: Neutralizing antibody against severe acute respiratory syndrome (SARS)‐coronavirus spike is highly effective for the protection of mice in the murine SARS model

doi: 10.1111/j.1348-0421.2008.00097.x

Figure Lengend Snippet: Concentrations of TNF‐α, IL‐6 and MCP‐1 in the lung lavage of vaccinated mice. Cytokines and chemokines in the lung lavage of vaccinated and control mice on day 3 after SARS‐CoV infection were assayed by flow cytometry using a mouse inflammatory cytokine (IL‐6, IL‐10, MCP‐1, IFN‐γ, TNF‐α and IL‐12p70) cytometric bead array kit. Bar shows the SEM ( n = 3).

Article Snippet: Cytokines and chemokines were assayed either using a mouse inflammatory cytokine (IL‐6, IL‐10, MCP‐1, IFN‐γ, TNF‐α and IL‐12p70) cytometric bead array kit (Becton Dickinson, San Jose, CA, USA) ( ) or using the Luminex 200 system (Luminex Co., Austin, TX, USA) as previously reported ( ).

Techniques: Control, Infection, Flow Cytometry

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Selective BET-bromodomain inhibition by JQ1 suppresses dendritic cell maturation and antigen-specific T-cell responses

doi: 10.1007/s00262-020-02665-x

Figure Lengend Snippet: JQ1 impairs activation and modulates cytokine production of murine BMDCs. BMDC were treated with JQ1 (filled square), respectively, DMSO (open square) on day 7 of culture overnight and challenged with LPS for 24 h. Immature DC (iDC) were DMSO-treated without final LPS challenge. a Relative RNA fold induction of surface molecules and chemokines, normalized to DMSO control. Triplicates of one experiment that stands representative for 3. b Expression of surface molecules after JQ1 treatment in varying doses [10, 50, 100, 250, and 500 nM) are shown. Pooling of three independent experiments. c Immunostaining of intracellular cytokines. Triplicates of one experiment. d Supernatant of cell culture was collected and analyzed for IL-6 production by ELISA. Pooling of three independent experiments

Article Snippet: IL-6 secretion in cell culture supernatant and mouse serum was measured using the OptEIA™ Mouse IL-6 Elisa Set (Becton Dickinson) following the manufacturer’s instructions.

Techniques: Activation Assay, Expressing, Immunostaining, Cell Culture, Enzyme-linked Immunosorbent Assay

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Selective BET-bromodomain inhibition by JQ1 suppresses dendritic cell maturation and antigen-specific T-cell responses

doi: 10.1007/s00262-020-02665-x

Figure Lengend Snippet: JQ1 reduces DC and T-cell immune response in vivo. C57BL/6 mice were treated with 25 mg/kg body weight JQ1 or vehicle for 4 days and challenged with 12.5 µg LPS daily on days 2–4. Analysis of splenic DCs and T cells was done on day 5. Each group was consistent of four individuals. a, b Exemplary histograms and proportion of CD86+, respectively, MHC II+ populations of splenic DCs. c Proportion of CD25+, CD44+, and CD62L- populations of splenic CD4+ T cells. d Proportion of CD25+, CD44+, and CD62L- populations of splenic CD8+ T cells. e Blood count of white and red blood cells. Blood was taken from tail veins. f Serum concentration of IL-6 measured by ELISA

Article Snippet: IL-6 secretion in cell culture supernatant and mouse serum was measured using the OptEIA™ Mouse IL-6 Elisa Set (Becton Dickinson) following the manufacturer’s instructions.

Techniques: In Vivo, Concentration Assay, Enzyme-linked Immunosorbent Assay

Journal: Cell Death & Disease

Article Title: Regulation of Zfp36 by ISGF3 and MK2 restricts the expression of inflammatory cytokines during necroptosis stimulation

doi: 10.1038/s41419-024-06964-4

Figure Lengend Snippet: WT and Ifnar1 −/− BMDMs were treated with LPS (1 ng/mL) in the presence of zVAD-fmk (50 µM) for 6 h and differential gene expression and GSEA was performed on microarray data comparing WT BMDMs treated with LPS+zVAD with Ifnar1 −/− BMDMs treated with LPS+zVAD ( A – D ). E Expression levels of TNFα, IFNβ, IL-6, and IL-10 were measured in the supernatants collected after treating WT and Ifnar1 −/− BMDMs for 7 h with LPS (1 ng/mL) and EMR (10 µM). F , G Western blot analysis was performed in cell extracts collected from WT and Ifnar1 −/− BMDMs at different time intervals following treatment with LPS (1 ng/mL) and EMR (10 µM). Each experiment was repeated at least three times. (**** P < 0.0001).

Article Snippet: Supernatants were collected from 96-well plates and the expression of cytokines was assessed using the mouse TNF-α (BD OptEIA #555268), mouse IL-6 (BD OptEIA #555240), mouse IL-10 (BD OptEIA #555252), mouse IL-12p70 (BD OptEIA #555256) and mouse IFN-β (R&D System #DY8234-05) according to the manufacturer’s instructions.

Techniques: Expressing, Microarray, Western Blot

Journal: Cell Death & Disease

Article Title: Regulation of Zfp36 by ISGF3 and MK2 restricts the expression of inflammatory cytokines during necroptosis stimulation

doi: 10.1038/s41419-024-06964-4

Figure Lengend Snippet: A Expression of TTP was analyzed by western blotting of cell extracts collected from WT and Ifnar1 −/− BMDMs at various time intervals following treatment with LPS (1 ng/mL) and/or EMR (10 µM). B – E Expression of TNFα, IFNβ, and IL-6 was measured in the supernatants collected at different time intervals after the treatment of WT and Zfp36 −/− BMDMs with LPS (1 ng/mL) ( B ), LPS (1 ng/mL)+EMR (10 µM) ( C ), IFNβ (10 ng/mL) ( D ), or IFNβ (10 ng/mL)+EMR (10 µM) ( E ). F , G Cell death was evaluated by staining of cells with Hoechst and PI at 24 h post treatment with LPS + /−EMR ( F , G ), IFNβ + /−EMR ( F, H ), TNFα ± EMR ( F , I ). Each experiment was repeated at least three times. (*** P < 0.001, **** P < 0.0001).

Article Snippet: Supernatants were collected from 96-well plates and the expression of cytokines was assessed using the mouse TNF-α (BD OptEIA #555268), mouse IL-6 (BD OptEIA #555240), mouse IL-10 (BD OptEIA #555252), mouse IL-12p70 (BD OptEIA #555256) and mouse IFN-β (R&D System #DY8234-05) according to the manufacturer’s instructions.

Techniques: Expressing, Western Blot, Staining

Journal: Cell Death & Disease

Article Title: Regulation of Zfp36 by ISGF3 and MK2 restricts the expression of inflammatory cytokines during necroptosis stimulation

doi: 10.1038/s41419-024-06964-4

Figure Lengend Snippet: A , B WT and Ifnar1 −/− BMDMs were stimulated with LPS (1 ng/mL), EMR (10 µM) and MK2 inhibitor (III, 5 µM), and the impact on TNFα secretion ( A ) and cell viability ( B ) was evaluated at 7 h and 24 h respectively. C – G WT, Ifnar1 −/− , Mk2 −/− , and Ifnar1 −/− Mk2 −/− BMDMs were stimulated with LPS (1 ng/mL)+EMR (10 µM). Cytokine expression was evaluated at various time intervals by ELISA ( C ). Activation of signaling proteins was evaluated by western blot analysis of cell extracts collected at various time intervals ( D ). Cell death was evaluated by staining cells with Hoechst and PI following stimulation of WT, Ifnar1 −/− , Mk2 −/− , and Ifnar1 −/− Mk2 −/− BMDMs with LPS (1 ng/mL)+EMR (10 µM), IFNβ (10 ng/mL)+EMR (10 µM), and TNFα (50 ng/mL)+EMR (10 µM) at 9 h ( F ) or 18 h ( E , G ). Each experiment was repeated at least three times. (* P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001).

Article Snippet: Supernatants were collected from 96-well plates and the expression of cytokines was assessed using the mouse TNF-α (BD OptEIA #555268), mouse IL-6 (BD OptEIA #555240), mouse IL-10 (BD OptEIA #555252), mouse IL-12p70 (BD OptEIA #555256) and mouse IFN-β (R&D System #DY8234-05) according to the manufacturer’s instructions.

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Activation Assay, Western Blot, Staining