Journal: Mediators of Inflammation

Article Title: SP600125 Attenuates Nicotine-Related Aortic Aneurysm Formation by Inhibiting Matrix Metalloproteinase Production and CC Chemokine-Mediated Macrophage Migration

doi: 10.1155/2016/9142425

Figure Lengend Snippet: Proposed mechanism of SP600125-mediated suppression of nicotine-related AAA in the C57BL/6J mouse model. AngII, angiotensin II; JNK, c-Jun N-terminal kinase; MCP-1, monocyte chemoattractant protein-1; RANTES, regulated-on-activation, normal T-cell expressed and secreted; MMP, matrix metalloproteinase; ECM, extracellular matrix.

Article Snippet: Slides were incubated with rabbit anti-mouse RANTES immunoglobulin G (IgG), rabbit anti-mouse MCP-1 IgG, rabbit anti-mouse MMP-2, rabbit anti-mouse MMP-9, or reagent-grade nonimmune rat IgG (R&D Systems, Minneapolis, MN, USA) overnight at 4°C in a humidified chamber.

Techniques: Activation Assay

Journal: Mediators of Inflammation

Article Title: SP600125 Attenuates Nicotine-Related Aortic Aneurysm Formation by Inhibiting Matrix Metalloproteinase Production and CC Chemokine-Mediated Macrophage Migration

doi: 10.1155/2016/9142425

Figure Lengend Snippet: SP600125 suppresses the protein expression of chemokines in aortic tissue. (a) Representative transverse sections of mouse aortic tissue obtained after transient perfusion with nicotine plus AngII, with or without SP600125. MCP-1 and RANTES proteins were undetectable in normal aorta but were abundant in nicotine plus AngII-induced AAA tissues (a1 and a2), where they appear to be expressed in the medial and outer smooth muscle cells. SP600125 inhibited the protein expression of MCP-1 and RANTES in AAA lesions (a3). (c, d) Representative western blots for MCP-1 and RANTES. The band optical density (OD) values (mean ± SD) of MCP-1 and RANTES were evaluated using ImageJ. GAPDH was used as an internal control and results are from independent triplicate experiments. Chemokine expression in the serum as detected by ELISA (e). ∗∗ P < 0.01 versus coadministration.

Article Snippet: Slides were incubated with rabbit anti-mouse RANTES immunoglobulin G (IgG), rabbit anti-mouse MCP-1 IgG, rabbit anti-mouse MMP-2, rabbit anti-mouse MMP-9, or reagent-grade nonimmune rat IgG (R&D Systems, Minneapolis, MN, USA) overnight at 4°C in a humidified chamber.

Techniques: Expressing, Western Blot, Control, Enzyme-linked Immunosorbent Assay

Journal: Mediators of Inflammation

Article Title: SP600125 Attenuates Nicotine-Related Aortic Aneurysm Formation by Inhibiting Matrix Metalloproteinase Production and CC Chemokine-Mediated Macrophage Migration

doi: 10.1155/2016/9142425

Figure Lengend Snippet: Expression of MCP-1 and RANTES under a concentration gradient of nicotine in MOVAS cells. Cellular mRNA expression levels of MCP-1 and RANTES (a, b) and the levels of secreted MCP-1 and RANTES in the supernatant (c, d) are shown. MCP-1 and RANTES expression as well as secretion was induced by nicotine in a dose-dependent fashion. The strongest expression was observed for 0.5 ng/mL and 5 ng/mL nicotine. Data are from independent triplicate experiments. ∗ P < 0.05 and ∗∗ P < 0.01 versus control; # P < 0.05 and ## P < 0.01 versus the group treated with 5 ng/mL nicotine.

Article Snippet: Slides were incubated with rabbit anti-mouse RANTES immunoglobulin G (IgG), rabbit anti-mouse MCP-1 IgG, rabbit anti-mouse MMP-2, rabbit anti-mouse MMP-9, or reagent-grade nonimmune rat IgG (R&D Systems, Minneapolis, MN, USA) overnight at 4°C in a humidified chamber.

Techniques: Expressing, Concentration Assay, Control

Journal: Mediators of Inflammation

Article Title: SP600125 Attenuates Nicotine-Related Aortic Aneurysm Formation by Inhibiting Matrix Metalloproteinase Production and CC Chemokine-Mediated Macrophage Migration

doi: 10.1155/2016/9142425

Figure Lengend Snippet: Influence of SP600125 on nicotine-induced expression of MCP-1 and RANTES in MOVAS cells. Cellular mRNA expression levels of MCP-1 and RANTES (a, b) and the levels of secreted MCP-1 and RANTES in the supernatant (c, d) are shown. Nicotine at 5 ng/mL could significantly upregulate the cellular mRNA expression as well as secretion of MCP-1 and RANTES, while SP600125 eliminated this effect. Data are from independent triplicate experiments. ∗ P < 0.05 and ∗∗ P < 0.01 versus control; # P < 0.05 and ## P < 0.01 versus the group treated with 5 ng/mL nicotine.

Article Snippet: Slides were incubated with rabbit anti-mouse RANTES immunoglobulin G (IgG), rabbit anti-mouse MCP-1 IgG, rabbit anti-mouse MMP-2, rabbit anti-mouse MMP-9, or reagent-grade nonimmune rat IgG (R&D Systems, Minneapolis, MN, USA) overnight at 4°C in a humidified chamber.

Techniques: Expressing, Control

Journal: Acta Neuropathologica Communications

Article Title: Brain injury drives optic glioma formation through neuron-glia signaling

doi: 10.1186/s40478-024-01735-w

Figure Lengend Snippet: a Following optic nerve crush (ON-CR) at 6 weeks of age, Nf1 flox/flox ; hGFAP-Cre mice at 12 weeks of age have increased numbers of CD3 + cells in their optic nerves relative to those receiving a sham operation ( n = 6); however, there was no change in b Ccl4 mRNA expression by qPCR ( n = 3). c Immediately after ON-CR, Nf1 flox/flox ; hGFAP-Cre mice received intraperitoneal injections of anti-CD3 (αCD3) antibodies every other day for 6 weeks. Control mice were injected with anti-IgG antibodies. Optic nerves were analyzed at 12 weeks of age. αCD3 antibody treatment (150 µg) reduced d CD3 + cell content ( n = 5) and e proliferation (%Ki67 + cells; n = 5). f αCD3 antibody treatment following ON-CR does not change Ccl5 RNA expression by qPCR compared to IgG-treated controls (150 µg; n = 3). Increased Ccl5 RNA expression in the optic nerves (qPCR) was observed 7 days after ON-CR in g Nf1 flox/flox ; hGFAP-Cre ( n = 3), h athymic ( Foxn1 nu/nu ; n = 4), and i Rag1 −/− mice ( n = 4) compared to sham controls. Scale bars: a , d , e 50 µm. Two-tailed Student’s t test (ns, not significant)

Article Snippet: Four-week-old Nf1 OPG mice were treated with 10 mg/kg NFκB inhibitor (NFκB-IN; Caffeic acid phenethyl ester, Fisher Scientific, 274310), 275 mg/kg PLX3397-containing or control chow pellets (Free Base), 1 mg/ml anti-IL-1β neutralizing antibodies (R&D System 1060-DE-100), anti-IgG2a control antibodies (R&D Systems), or memantine hydrochloride (20 mg/kg) by intraperitoneal injection.

Techniques: Expressing, Control, Injection, RNA Expression, Two Tailed Test

Journal: Acta Neuropathologica Communications

Article Title: Brain injury drives optic glioma formation through neuron-glia signaling

doi: 10.1186/s40478-024-01735-w

Figure Lengend Snippet: a Top 3 identified pathways enriched in the optic nerves of Nf1 flox/flox ; hGFAP-Cre mice after optic nerve crush (ON-CR) relative to sham controls following filtering of differentially expressed transcripts from bulk RNA sequencing. b Increased IL-1β RNA expression (qPCR) is observed in the optic nerves of 12-week-old Nf1 flox/flox ; hGFAP-Cre mice following ON-CR at 6 weeks of age relative to sham controls ( n = 4). c Representative IL-1β immunostaining in the optic nerves of Nf1 flox/flox ; hGFAP-Cre mice following ON-CR relative to sham control mice. d RNAscope (in situ RNA hybridization) demonstrates that oligodendrocytes (Olig2 + cells) express IL-1β. e Immunostaining reveals that the majority of the Olig2 + cells co-label with CC1, but not with NG2. f Increasing IL-1β concentrations (0–75 ng/ml) increases microglia Ccl5 protein expression in vitro. g IL-1β-induced microglial Ccl5 production is attenuated by 10 mg/kg NFκB inhibitor (NFκB-IN; Caffeic acid phenethyl ester) treatment in vitro ( n = 4). h Anti-IL1β neutralizing antibody treatment (1 mg/ml) of Nf1 flox/flox ; hGFAP-Cre mice immediately after ON-CR at 6 weeks of age results in decreased i optic nerve proliferation (%Ki67 + cells; n = 5) and j Ccl5 mRNA expression (qPCR; n = 3) when analyzed at 12 weeks of age compared to IgG-treated (1 mg/ml) controls. Data are presented as the means ± SEM. Scale bars: c , d , e , i 50 µm. b , c , e , i , j . Two-tailed Student’s t test; f , g One-way ANOVA with Bonferroni post-test correction

Article Snippet: Four-week-old Nf1 OPG mice were treated with 10 mg/kg NFκB inhibitor (NFκB-IN; Caffeic acid phenethyl ester, Fisher Scientific, 274310), 275 mg/kg PLX3397-containing or control chow pellets (Free Base), 1 mg/ml anti-IL-1β neutralizing antibodies (R&D System 1060-DE-100), anti-IgG2a control antibodies (R&D Systems), or memantine hydrochloride (20 mg/kg) by intraperitoneal injection.

Techniques: RNA Sequencing, RNA Expression, Immunostaining, Control, RNAscope, In Situ, Hybridization, Expressing, In Vitro, Two Tailed Test

Journal: Acta Neuropathologica Communications

Article Title: Brain injury drives optic glioma formation through neuron-glia signaling

doi: 10.1186/s40478-024-01735-w

Figure Lengend Snippet: a Traumatic brain injury (TBI) in Nf1 flox/flox ; hGFAP-Cre mice at 6 weeks of age results in increased b optic nerve volume ( n = 6) and c proliferation (%Ki67 + cells; n = 7) relative to sham treated mice when analyzed at 12 weeks of age, as well as increased TAMs (%Iba1 + cells) and CD3 + T cell content. Increased d Ccl5 RNA expression (qPCR) and e glutamate levels are observed in the optic nerves of Nf1 flox/flox ; hGFAP-Cre mice ( n = 4) 7 days after TBI compared to sham controls. f αIL-1β neutralizing antibody (1 mg/ml) and h memantine treatment (20 mg/kg) both decrease proliferation (%Ki67 + cells; n = 5) and g , i Ccl5 expression ( n = 3) following TBI relative to their respective control mice (IgG and vehicle treatment groups). Data are presented as the means ± SEM. Scale bar: b 100 μm; c , f , i 50 μm. Two-tailed Student’s t test

Article Snippet: Four-week-old Nf1 OPG mice were treated with 10 mg/kg NFκB inhibitor (NFκB-IN; Caffeic acid phenethyl ester, Fisher Scientific, 274310), 275 mg/kg PLX3397-containing or control chow pellets (Free Base), 1 mg/ml anti-IL-1β neutralizing antibodies (R&D System 1060-DE-100), anti-IgG2a control antibodies (R&D Systems), or memantine hydrochloride (20 mg/kg) by intraperitoneal injection.

Techniques: RNA Expression, Expressing, Control, Two Tailed Test

Journal: Vaccines

Article Title: Simultaneous Administration of Recombinant Measles Viruses Expressing Respiratory Syncytial Virus Fusion (F) and Nucleo (N) Proteins Induced Humoral and Cellular Immune Responses in Cotton Rats

doi: 10.3390/vaccines7010027

Figure Lengend Snippet: The proportion of CD8 + /IFN-γ + cells in spleen cells obtained four days after the RSV challenge (Day 88), stimulated with UV-inactivated RSV Long strain, F, NP, and M2-1 peptides. Spleen cells were obtained after the RSV challenge and analyzed for IFN-γ production by CD8 + cells. The bar represents the mean of 3~4 rats per group ± SEM. (* p < 0.05).

Article Snippet: After the stimulation, splenocytes were washed and incubated with an anti-CD8 antibody (R&D Systems, Minneapolis, MN, USA) at 4 °C for 30 min. Splenocytes were fixed with fixation buffer in the Cytofix/cytoperm kit, and intracellular cytokines were stained with a goat IgG antibody against IFN-γ (R&D Systems, USA) and anti-goat IgG PE-Cy7 (Santa Cruz Biotechnology, Inc., Dallas, TX, USA) at 4 °C for 60 min. Cellular populations were analyzed using flow cytometry with Cytomics FC 500 (Beckman Coulter, Inc., Indianapolis, IN, USA) and counted until 100,000 cells.

Techniques:

Journal: Vaccines

Article Title: Simultaneous Administration of Recombinant Measles Viruses Expressing Respiratory Syncytial Virus Fusion (F) and Nucleo (N) Proteins Induced Humoral and Cellular Immune Responses in Cotton Rats

doi: 10.3390/vaccines7010027

Figure Lengend Snippet: Detection of IFN-γ + expressing cells in lung homogenates. Lung homogenates were obtained after the RSV challenge on Day 88. IFN-γ-producing cells were separated further into CD8 + and CD4 + cells. The bar represents the mean of 3~4 rats per group ± SEM. (* p < 0.05).

Article Snippet: After the stimulation, splenocytes were washed and incubated with an anti-CD8 antibody (R&D Systems, Minneapolis, MN, USA) at 4 °C for 30 min. Splenocytes were fixed with fixation buffer in the Cytofix/cytoperm kit, and intracellular cytokines were stained with a goat IgG antibody against IFN-γ (R&D Systems, USA) and anti-goat IgG PE-Cy7 (Santa Cruz Biotechnology, Inc., Dallas, TX, USA) at 4 °C for 60 min. Cellular populations were analyzed using flow cytometry with Cytomics FC 500 (Beckman Coulter, Inc., Indianapolis, IN, USA) and counted until 100,000 cells.

Techniques: Expressing