Journal: Scientific Reports

Article Title: High-intensity interval training remodels perineuronal nets in the medial prefrontal cortex to drive microglial polarization and alleviate osteoarthritis pain

doi: 10.1038/s41598-026-40823-w

Figure Lengend Snippet: HIIT reduced PNN accumulation and promoted microglial M2 polarization in the mPFC. (A and B) Triple immunofluorescence staining in the mPFC showing PNN, Iba1, and iNOS ( A ) or PNN, Iba1, and Arg1 ( B ). OA rats showed increased PNN expression and pro-inflammatory (Iba1 + iNOS⁺) microglia, while HIIT reduced PNN levels and promoted anti-inflammatory (Iba1 + Arg1⁺) phenotypes. (C and D) Protein expression of iNOS, Arg1, IL-10, IL-1β, and TNF-α in the mPFC, detected by western blot ( C ) and quantified by bar graphs ( D ) ( n = 6). Statistical methods used: One-way ANOVA. ** p < 0.01 vs. the sham group, ## p < 0.01 vs. the OA group. Data are represented as mean ± SD.

Article Snippet: Equal amounts of protein (30–50 μg) were separated on 10–12% SDS-PAGE gels (FuturePAGE, ACE Biotechnology, ET15420LGel), transferred to PVDF membranes (Millipore, Billerica, MA, USA, IPVH00010), blocked with 5% skim milk (BD Biosciences, Franklin Lakes, NJ, USA) for 1 h, and incubated overnight at 4 °C with primary antibodies against β-actin (1:5000, Servicebio, GB15003-100), COL2A1 (1:1000, Santa Cruz Biotechnology, sc-52658), MMP13 (1:1000, Proteintech, 18165-1-AP), iNOS (1:1000, Santa Cruz Biotechnology, sc-7271), Arg1 (1:1000, Santa Cruz Biotechnology, sc-47715), IL-10 (1:1000, Santa Cruz Biotechnology, sc-32815), IL-1β (1:1000, Santa Cruz Biotechnology, sc-12742), and TNF-α (1:1000, Santa Cruz Biotechnology, sc-52746).

Techniques: Immunofluorescence, Staining, Expressing, Western Blot

Journal: Scientific Reports

Article Title: High-intensity interval training remodels perineuronal nets in the medial prefrontal cortex to drive microglial polarization and alleviate osteoarthritis pain

doi: 10.1038/s41598-026-40823-w

Figure Lengend Snippet: PNNs degradation and HIIT alleviate neuroinflammation in the mPFC of OA rats. ( A - D ) Representative triple immunofluorescence staining images of the mPFC, showing the distribution patterns of PNNs, Iba1 (microglial marker), and iNOS ( A ) or Arg1 ( C ). Quantification of panel A is shown in ( B ), and quantification of panel C is shown in ( D ) ( n = 6). ( E and F ) Western blot analysis and corresponding quantification of inflammatory markers, including iNOS, Arg1, IL-1β, TNF-α, and IL-10 in mPFC tissue lysates ( n = 6). Statistical methods used: One-way ANOVA. ** p < 0.01 vs. the OA+Vehicle group, ns not significant vs. the OA+ChABC group.Data are presented as mean ± SD.

Article Snippet: Equal amounts of protein (30–50 μg) were separated on 10–12% SDS-PAGE gels (FuturePAGE, ACE Biotechnology, ET15420LGel), transferred to PVDF membranes (Millipore, Billerica, MA, USA, IPVH00010), blocked with 5% skim milk (BD Biosciences, Franklin Lakes, NJ, USA) for 1 h, and incubated overnight at 4 °C with primary antibodies against β-actin (1:5000, Servicebio, GB15003-100), COL2A1 (1:1000, Santa Cruz Biotechnology, sc-52658), MMP13 (1:1000, Proteintech, 18165-1-AP), iNOS (1:1000, Santa Cruz Biotechnology, sc-7271), Arg1 (1:1000, Santa Cruz Biotechnology, sc-47715), IL-10 (1:1000, Santa Cruz Biotechnology, sc-32815), IL-1β (1:1000, Santa Cruz Biotechnology, sc-12742), and TNF-α (1:1000, Santa Cruz Biotechnology, sc-52746).

Techniques: Immunofluorescence, Staining, Marker, Western Blot

Journal: Scientific Reports

Article Title: High-intensity interval training remodels perineuronal nets in the medial prefrontal cortex to drive microglial polarization and alleviate osteoarthritis pain

doi: 10.1038/s41598-026-40823-w

Figure Lengend Snippet: PNN remodeling precedes microglial polarization. ( A - D ) Representative triple immunofluorescence staining images of the mPFC, showing PNN, Iba1, and iNOS ( A ), or PNN, Iba1, and Arg1 ( C ). Quantification of panel A is shown in ( B ), and quantification of panel C is shown in ( D ) ( n = 6). Statistical methods used: One-way ANOVA. ** p < 0.01. Data are presented as mean ± SD.

Article Snippet: Equal amounts of protein (30–50 μg) were separated on 10–12% SDS-PAGE gels (FuturePAGE, ACE Biotechnology, ET15420LGel), transferred to PVDF membranes (Millipore, Billerica, MA, USA, IPVH00010), blocked with 5% skim milk (BD Biosciences, Franklin Lakes, NJ, USA) for 1 h, and incubated overnight at 4 °C with primary antibodies against β-actin (1:5000, Servicebio, GB15003-100), COL2A1 (1:1000, Santa Cruz Biotechnology, sc-52658), MMP13 (1:1000, Proteintech, 18165-1-AP), iNOS (1:1000, Santa Cruz Biotechnology, sc-7271), Arg1 (1:1000, Santa Cruz Biotechnology, sc-47715), IL-10 (1:1000, Santa Cruz Biotechnology, sc-32815), IL-1β (1:1000, Santa Cruz Biotechnology, sc-12742), and TNF-α (1:1000, Santa Cruz Biotechnology, sc-52746).

Techniques: Immunofluorescence, Staining

Journal: Italian Journal of Animal Science

Article Title: Protective effect of chitosan oligosaccharide against oxidative damage of peripheral blood mononuclear cells in dairy cows induced by diethylenetriamine/nitric oxide via NF-κB signalling pathway

doi: 10.1080/1828051x.2020.1772131

Figure Lengend Snippet: Figure 2. Effects of chitosan oligosaccharideon on the phosphorylation level of NF-kB pathway, the protein expression level of nitric oxide synthase (iNOS) and interleukin-1b (IL-1b). Expressions of IL-1b(A); phosphorylated IjB kinase b (P-IKKb) (B); phosphorylated inhibitor of nuclear factor kappa-Ba (P-IjBa) (C); phosphorylated nuclear factor kappa-Bp65(P-NF-jBp65) (D); iNOS (E), protein levels were detected by western blotting and normalised to beta-actin (b-Actin) levels. CTR¼ control treatment, without chitosan oligosacchar- ides addition; COS40, COS80, COS160, and COS320¼ treated with 40, 80, 160, and 320lg/mL chitosan oligosaccharides respectively.

Article Snippet: The membranes were incubated in the following primary antibodies: primary polyclonal rabbit anti-iNOS antibody (NBP1-97471, Novus Biological, USA), rabbit anti-IL-1b (Cell Signalling Technology, Boston, MA), anti-IKKb, monoclonal rat anti-IjBa, monoclonal rabbit anti-NF-jBp65 antibody(NF-jB pathway Sampler Kit 9963,CST, MA), anti-phospho-IKKb, anti-phospho-IjBa, and anti-phospho-NF-jBp65 (NF-jB pathway Sampler Kit 9963, CST, MA).

Techniques: Phospho-proteomics, Expressing, Western Blot, Control

Journal: Frontiers in Immunology

Article Title: ATF3 Stimulates IL-17A by Regulating Intracellular Ca 2+ /ROS-Dependent IL-1β Activation During Streptococcus pneumoniae Infection

doi: 10.3389/fimmu.2018.01954

Figure Lengend Snippet: ATF3 suppresses ROS generation and iNOS expression. (A) Fold change in ROS levels at 0, 2, 4, and 6 hpi compared to that in non-infected WT cells. (B) iNOS expression and (C) IHC (anti-iNOS) at 0 and 6 hpi ( n = 4). * P < 0.05, ** P < 0.01, two-way analysis of variance.

Article Snippet: The membranes were blocked with skim milk for 2 h, and then probed overnight with anti-GBP5 (gtx31537, Gentex, Zeeland, MI, USA), anti-iNOS (NB300-605, Novus International, St. Louis, MO, USA), anti-IL-1β (AF-401-SP, R&D Systems, Minneapolis, MN, USA), or anti-β-actin (sc-47778, Santa Cruz Biotechnology, Dallas, TX, USA).

Techniques: Expressing, Infection

Journal: Frontiers in Immunology

Article Title: ATF3 Stimulates IL-17A by Regulating Intracellular Ca 2+ /ROS-Dependent IL-1β Activation During Streptococcus pneumoniae Infection

doi: 10.3389/fimmu.2018.01954

Figure Lengend Snippet: ATF3 plays an important role in IL-17A production in response to S. pneumoniae infection. Lung macrophages rapidly phagocytose invading S. pneumoniae during infection, resulting in ROS production, ER stress, and ATF3 activation. ATF3 then inhibits ROS-induced iNOS expression to promote NLRP3 inflammasome and GBP5 activation, triggering IL-1β secretion and the subsequent activation of γδ T cells in the lung .

Article Snippet: The membranes were blocked with skim milk for 2 h, and then probed overnight with anti-GBP5 (gtx31537, Gentex, Zeeland, MI, USA), anti-iNOS (NB300-605, Novus International, St. Louis, MO, USA), anti-IL-1β (AF-401-SP, R&D Systems, Minneapolis, MN, USA), or anti-β-actin (sc-47778, Santa Cruz Biotechnology, Dallas, TX, USA).

Techniques: Infection, Activation Assay, Expressing

Journal: Acta Pharmacologica Sinica

Article Title: Efficacy of subantimicrobial-dose doxycycline against nitrosative stress in chronic periodontitis

doi: 10.1038/aps.2012.129

Figure Lengend Snippet: Photomicrographs of representative iNOS-positive and 3-nitrotyrosine-positive stains in gingival tissue from patients with chronic periodontitis (immunohistochemical staining): (A) countless iNOS+ PMNs and monocytes (scale bar, 50 μm); (B) decreased iNOS+ cells in the SRP+placebo group (scale bar, 50 μm); (C) sparse iNOS+ cells in SRP+SDD group (scale bar, 50 μm); (D) countless 3NT+ PMNs and monocytes (scale bar, 100 μm); (E) decreased 3NT+ cells in the SRP+placebo group (scale bar, 50 μm); (F) sparse 3NT+ cells in the SRP+SDD group (scale bar, 50 μm). This figure is representative of 3 measurements (×400). iNOS+=induced nitric oxide synthase positive cells; 3NT+=3-nitrotyrosine positive cells; SRP=scaling and root planing; SDD=subantimicrobial dose of doxycycline.

Article Snippet: The following primary monoclonal antibodies were used: anti iNOS (R&D Systems, RD-MAB9502, MN, USA), anti 3-nitrotyrosine (3NT) (Novus Europe, KA0445, ABNOVA, Cambridge, UK).

Techniques: Immunohistochemical staining, Staining

Journal: Acta Pharmacologica Sinica

Article Title: Efficacy of subantimicrobial-dose doxycycline against nitrosative stress in chronic periodontitis

doi: 10.1038/aps.2012.129

Figure Lengend Snippet: Periodontal immunohystochemical and serum nitrosative stress parameters of the patients.

Article Snippet: The following primary monoclonal antibodies were used: anti iNOS (R&D Systems, RD-MAB9502, MN, USA), anti 3-nitrotyrosine (3NT) (Novus Europe, KA0445, ABNOVA, Cambridge, UK).

Techniques: