Journal: Immunity, Inflammation and Disease

Article Title: Inhibition of TMUB1 blocks apoptosis and NF‐κB pathway‐mediated inflammation in recurrent spontaneous abortion

doi: 10.1002/iid3.879

Figure Lengend Snippet: Effects of LPS on apoptosis and inflammation in placental tissues of pregnant mice (A) Representative images of TUNEL staining at magnification X200 (Scale bars: 100 µm) and magnification X400 (Scale bars: 50 µm). (B) The mRNA levels of IL‐6 and TNF‐α were measured with quantitative RT‐PCR. (C) The content of IL‐6 and TNF‐α was quantified by ELISA. (D) Western blot analysis for p‐p65 and p65 (left), and the quantification of the p‐p65/p65 ratio (right). ELISA, enzyme‐linked immunosorbent assay; IL‐6, interleukin‐6; LPS, lipopolysaccharide; mRNA, messenger RNA; RT‐PCR, real‐time polymerase chain reaction; TNF‐α, tumor necrosis factor‐α; TUNEL, terminal deoxynucleotidyl transferase dUTP nick end labeling.

Article Snippet: The content of IL‐6 and TNF‐α was measured using the Mouse TNF‐α ELISA Kit, Mouse IL‐6 ELISA Kit, Human TNF‐α ELISA Kit, and Human IL‐6 ELISA Kit (LIANKE Biotech) according to the manufacturer's instructions.

Techniques: TUNEL Assay, Staining, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Western Blot, Reverse Transcription Polymerase Chain Reaction, Real-time Polymerase Chain Reaction

Journal: Immunity, Inflammation and Disease

Article Title: Inhibition of TMUB1 blocks apoptosis and NF‐κB pathway‐mediated inflammation in recurrent spontaneous abortion

doi: 10.1002/iid3.879

Figure Lengend Snippet: Effects of TMUB1 silencing on inflammation in LPS‐induced trophoblast cells. (B) The levels of IL‐6 and TNF‐α in the different treatments were measured by quantitative RT‐PCR and ELISA. (B) The expression of p‐IKKα/β and IKKα was detected by western blot, and the ratio of p‐IKKα/β/IKKα was quantified in different groups. (C) The expression of p‐p65 and p65 was detected by western blot, and the ratio of p‐p65/p65 was quantified in different groups. (D) The protein content of NF‐κB p65 in the cytoplasm and nucleus was detected by western blot. ELISA, enzyme‐linked immunosorbent assay; IL‐6, interleukin‐6; LPS, lipopolysaccharide; RT‐PCR, real‐time polymerase chain reaction; TMUB1, transmembrane and ubiquitin‐like domain containing 1; TNF‐α, tumor necrosis factor‐α.

Article Snippet: The content of IL‐6 and TNF‐α was measured using the Mouse TNF‐α ELISA Kit, Mouse IL‐6 ELISA Kit, Human TNF‐α ELISA Kit, and Human IL‐6 ELISA Kit (LIANKE Biotech) according to the manufacturer's instructions.

Techniques: Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Expressing, Western Blot, Reverse Transcription Polymerase Chain Reaction, Real-time Polymerase Chain Reaction, Ubiquitin Proteomics

Journal: Immunity, Inflammation and Disease

Article Title: Inhibition of TMUB1 blocks apoptosis and NF‐κB pathway‐mediated inflammation in recurrent spontaneous abortion

doi: 10.1002/iid3.879

Figure Lengend Snippet: Effects of TMUB1 silencing in LPS‐induced mice. Pregnant mice were injected with lentivirus knocking down TMUB1 and then injected intraperitoneally with 0.15 μg/g LPS to induce abortion. (A) Pathological and anatomical observation of uteruses in pregnant mice. (B) The embryo absorption rate and abortion rate were calculated respectively in pregnant mice. (C) TUNEL staining was used to detect apoptosis in LPS‐induced mice with TMUB1 knockdown. Scale bar: 50 µm. (D) The mRNA levels of IL‐6 and TNF‐α were measured by quantitative RT‐PCR. (E) The protein content of p‐p65 and p65 was detected by western blot, and the ratio of p‐p65/p65 was quantified in different groups. DAPI, 4′,6‐diamidino‐2‐phenylindole; LPS, lipopolysaccharide; mRNA, messenger; RT‐PCR, real‐time polymerase chain reaction; TMUB1, transmembrane and ubiquitin‐like domain containing 1; TUNEL, terminal deoxynucleotidyl transferase dUTP nick end labeling.

Article Snippet: The content of IL‐6 and TNF‐α was measured using the Mouse TNF‐α ELISA Kit, Mouse IL‐6 ELISA Kit, Human TNF‐α ELISA Kit, and Human IL‐6 ELISA Kit (LIANKE Biotech) according to the manufacturer's instructions.

Techniques: Injection, TUNEL Assay, Staining, Knockdown, Quantitative RT-PCR, Western Blot, Reverse Transcription Polymerase Chain Reaction, Real-time Polymerase Chain Reaction, Ubiquitin Proteomics

Journal: Journal of Inflammation Research

Article Title: Tumor-Infiltrating PD-L1+ Neutrophils Induced by GM-CSF Suppress T Cell Function in Laryngeal Squamous Cell Carcinoma and Predict Unfavorable Prognosis

doi: 10.2147/JIR.S347777

Figure Lengend Snippet: LSCC tumor microenvironment remodels and regulates the phenotype and function of neutrophils. ( A ) TCCS or cultural medium from AMC-HN-8 cells upregulated the level of immunosuppressive molecule PD-L1 on neutrophils. ( B ) ELISA detected the concentration of GM-CSF, G-CSF and TGF-β1 in the plasma, TCCS and NTCCS of LSCC patients. ( C ) Representative images were shown for GM-CSF IHC staining (magnification, X 50) of tumor tissues and normal tissues from LSCC patients. Bars = 500μm. ( D ) Cytokine GM-CSF most significantly upregulated the level of immunosuppressive molecule PD-L1 on neutrophils. ( E-G ) Representative histograms and statistics analysis of the proliferative capacity of CD4+T cells and CD8+ T cells following the coculture of PANs from LSCC patients with PBMCs at different stimuli. ( H and I ) IFN-γ and TNF-α production of T cells following the coculture of PANs from LSCC patients with PBMCs at different stimuli. ns = p > 0.05, *p < 0.05, **p < 0.01, ***p < 0.001.

Article Snippet: TNF-α and INF-γ release in culture supernatants of lymphocytes proliferation assay were detected by ELISA kits (70-EK1821, 70-EK1801, MultiSciences, Hangzhou, China).

Techniques: Enzyme-linked Immunosorbent Assay, Concentration Assay, Clinical Proteomics, Immunohistochemistry

Journal: Brazilian Journal of Medical and Biological Research

Article Title: L-carnitine reduces acute lung injury via mitochondria modulation and inflammation control in pulmonary macrophages

doi: 10.1590/1414-431X2023e12830

Figure Lengend Snippet: Lipid accumulation in pulmonary macrophages increased in a lipopolysaccharide (LPS)-induced acute lung injury (ALI) murine model. A and B , LPS administration dramatically increased total cell number and neutrophil and lymphocyte counts ( A ), as well as the chemokine keratinocyte chemoattractant (KC) and cytokines interleukin (IL)-6, tumor necrosis factor (TNF)-α, and IL-1β ( B ) levels in the mouse bronchoalveolar lavage fluid (BALF). C , Representative images of lung sections stained with HE (left panels), myeloperoxidase (MPO) (middle panels), and Oil Red O (right four panels) from mice with or without LPS challenge. Red arrows indicate macrophages. Scale bar in the left six panels: 50 μm; scale bar in the two far right panels: 20 μm. Data are reported as means±SE (n=5-6). *P<0.05, **P<0.01, ***P<0.001. Statistical analysis was performed using two-tailed unpaired Student t -test.

Article Snippet: The levels of human interleukin-6 (IL-6) (MultiSciences Biotech, EK106/2-96, China), tumor necrosis factor-α (TNF-α) (MultiSciences Biotech, EK182-96), and IL-1β (MultiSciences Biotech, EK101B-96) in the cell supernatants of the 96-well plate were detected by ELISA kits according to the manufacturer's instructions.

Techniques: Staining, Two Tailed Test

Journal: Brazilian Journal of Medical and Biological Research

Article Title: L-carnitine reduces acute lung injury via mitochondria modulation and inflammation control in pulmonary macrophages

doi: 10.1590/1414-431X2023e12830

Figure Lengend Snippet: L-carnitine (Lca) suppressed lipopolysaccharide (LPS)-induced inflammation in THP-1-derived macrophages. A and B , Effects of Lca alone or in combination with LPS on cell viability of THP-1 cell-derived macrophages. C and D , Effects of Lca on LPS-elevated interleukin (IL)-6, tumor necrosis factor (TNF)-α, IL-17A mRNA ( C ), and IL-6, TNF-α, IL-1β secretion ( D ) in THP-1 cell-derived macrophages. E , Immunoblotting and related quantitative analysis of p-p65 and NLRP3 level for pro-inflammatory pathway activation in each group. Data are reported as means±SE (n=3-4). *P<0.05, **P<0.01, ***P<0.001, ns: not significant ( A - D ). *P<0.05, **P<0.01 vs LPS group ( E ). Statistical analysis was performed using one-way ANOVA followed by Bonferroni's post hoc test.

Article Snippet: The levels of human interleukin-6 (IL-6) (MultiSciences Biotech, EK106/2-96, China), tumor necrosis factor-α (TNF-α) (MultiSciences Biotech, EK182-96), and IL-1β (MultiSciences Biotech, EK101B-96) in the cell supernatants of the 96-well plate were detected by ELISA kits according to the manufacturer's instructions.

Techniques: Derivative Assay, Western Blot, Activation Assay