hmgb1  (Novus Biologicals)

Journal: European Journal of Clinical Investigation

Article Title: FNDC4 and FNDC5 Attenuate SARS‐CoV‐2 S1‐Induced Inflammatory Responses in Human Adipose Tissue

doi: 10.1111/eci.70215

Figure Lengend Snippet: Effect of FNDC4 and FNDC5 on SARS‐CoV‐2 spike protein S1 subunit‐mediated macrophage M1 polarisation and crosstalk with adipocytes. Effect of co‐incubation of SARS‐CoV‐2 spike protein S1 subunit (100 ng/mL) with FNDC4 (10 ng/mL) or FNDC5 (10 ng/mL) on gene expression of several factors involved in the polarisation of human THP‐1 macrophages toward M1 ( NOS2 ) or M2 ( ARG1 ) phenotypes. as well as in the production of inflammatory ( IL1B ) and anti‐inflammatory ( IL10 ) cytokines in the absence (A) or presence (B) of adipocyte‐conditioned medium (ACM 40%) derived from adipocyte cultures from patients with obesity ( n = 10 per group). Gene expression in unstimulated THP‐1 M0 macrophages was assumed to be 1. (C) HMGB1 release to the culture media in macrophages exposed to ACM 40% in the presence of S1 subunit with FNDC4 or FNDC5. Statistical differences were analysed by two‐way ANOVA followed by a post hoc Tukey's test. a p < 0.05 effect of the SARS‐CoV‐2 S1 subunit; b p < 0.05 effect of treatment with FNDC4 or FNDC5.

Article Snippet: The concentrations of HMGB1 (NBP2‐62766, Novus Biologicals, Cambridge, UK) released to the MCM were assessed by commercially available ELISA kits according to the manufacturer's protocols.

Techniques: Incubation, Gene Expression, Derivative Assay

Journal: European Journal of Clinical Investigation

Article Title: FNDC4 and FNDC5 Attenuate SARS‐CoV‐2 S1‐Induced Inflammatory Responses in Human Adipose Tissue

doi: 10.1111/eci.70215

Figure Lengend Snippet: SARS‐CoV‐2 spike protein S1 subunit promotes a proinflammatory phenotype in visceral adipocytes. (A) Effect of co‐incubation of SARS‐CoV‐2 spike protein S1 subunit (100 ng/mL) with FNDC4 (10 ng/mL) or FNDC5 (10 ng/mL) on protein expression of adiponectin and HMGB1 expression in human visceral adipocytes ( n = 6 per group). (B) HMGB1 gene expression in FNDC4 ‐ and FNDC5 ‐knockdown adipocytes ( n = 8 per group). Statistical differences were analysed by two‐way ANOVA or one‐way followed by a post hoc Tukey's test in case of interaction. * p < 0.05 versus unstimulated or siRNA control cells; † p < 0.05 versus SARS‐CoV‐2 S1‐treated cells.

Article Snippet: The concentrations of HMGB1 (NBP2‐62766, Novus Biologicals, Cambridge, UK) released to the MCM were assessed by commercially available ELISA kits according to the manufacturer's protocols.

Techniques: Incubation, Expressing, Gene Expression, Knockdown, Control

Journal: Journal of Inflammation Research

Article Title: Cell Death Index Predicts Sepsis Outcomes and Highlights Necroptosis as a Therapeutic Target

doi: 10.2147/JIR.S577930

Figure Lengend Snippet: Necrostatin-1 significantly improved sepsis outcomes in CLP models. (a) Serum levels of HMGB1 and CASP8 in clinical patients. (b) Schematic of the animal experimental design. (c) Survival rates of CLP mice with or without Nec-1 (10 mg/kg). (d) The mRNA levels of PDZD8 and ADRB2 were analyzed by RT-PCR in the peripheral blood of mice. (e) At the 22h post-CLP, the levels of inflammatory cytokines (TNF-α and IL-6) in plasma were detected by ELISA tests. (f) Hematoxylin–eosin staining of lung, liver, kidney and spleen sections, with red arrows indicating pathological areas. Scale bar = 50 μm. All data were expressed as mean ± SEM (n=3). *P< 0.05, **P< 0.01, ***P< 0.001.

Article Snippet: According to the manufacturer’s protocols, concentrations of HMGB1 (Elabscience, E-EL-H1554, Wuhan, China), CASP8 (Elabscience, E-EL-H0659, Wuhan, China), IL-6 (Solarbio, SEKM-0034, Beijing China), and TNF-α (ABclonal, RK04595, Wuhan, China) were measured using ELISA kits with standard curves ( Table S6 and 7 ).

Techniques: Reverse Transcription Polymerase Chain Reaction, Clinical Proteomics, Enzyme-linked Immunosorbent Assay, Staining

Journal: Journal of Extracellular Vesicles

Article Title: Chemoradiotherapy‐Integrated Tumor Cell‐Derived Microparticles Mediate Tumor Eradication in Malignant Pleural Effusion

doi: 10.1002/jev2.70277

Figure Lengend Snippet: CR‐MPs promote APCs activation in vitro . (A) MFI of CRT on control, MTX treated, RT‐MPs treated and CR‐MPs@MTX treated LLC cells. (B, C) Relative (B) HMGB1 level and (C) ATP level in culture supernatant of control, MTX treated, RT‐MPs treated and CR‐MPs@MTX treated LLC cells. (D) Representative immunofluorescence images of CRT (red) on control, MTX treated, RT‐MPs treated and CR‐MPs@MTX treated H1299 cells. Scale bar = 20 µm. (E, F) MFI of (E) CD80 and (F) CD206 on BMDMs co‐cultured with control, MTX treated, RT‐MPs treated and CR‐MPs@MTX treated LLC cells. (G) Phagocytosis ratio of BMDMs to control, MTX treated, RT‐MPs treated and CR‐MPs@MTX treated LLC cells. (H) Uptake of PKH26‐labeled, LLC‐derived CR‐MPs@MTX by BMDCs was assessed by flow cytometry at 4, 12, and 24 h. (I, J) MFI of (I) CD80 and (J) CD86 on control BMDCs and BMDCs treated with MTX, RT‐MPs and CR‐MPs@MTX. (K) KEGG pathway enrichment analysis in CR‐MPs@MTX‐treated BMDCs versus control BMDCs. (L) GSEA of the interferon‐alpha (IFNα) response pathway in CR‐MPs@MTX‐treated BMDCs versus control BMDCs. (M) Western blot analysis of phospho‐TBK1, phospho‐p65, phospho‐IRF3, and actin in CR‐MPs@MTX treated or MTX treated BMDCs at 5‐, 10‐, 30‐, and 240‐minute post‐treatment. (N) RT‐qPCR evaluated expression of indicated genes in BMDCs after 24‐hour treatment with MTX, RT‐MPs, or CR‐MPs@MTX, compared with untreated control. All data are presented as mean ± s.d. One‐way ANOVA with Tukey's multiple comparisons test was performed. * P < 0.05, ** P < 0.01, and *** P < 0.001; ns, no significance.

Article Snippet: Mouse HMGB1 (EM0382, Finetest) and human HMGB1 (E‐EL‐H1554, Elabscience) produced in the supernatants were evaluated by ELISA assays according to the manufacturer's instructions.

Techniques: Activation Assay, In Vitro, Control, Immunofluorescence, Cell Culture, Labeling, Derivative Assay, Flow Cytometry, Western Blot, Quantitative RT-PCR, Expressing

Journal: eBioMedicine

Article Title: Cytokine mRNA-based therapy alleviates dendritic cell and T cell paucity to eliminate aggressive pancreatic cancer in preclinical mouse models

doi: 10.1016/j.ebiom.2026.106137

Figure Lengend Snippet: Enhancement of immunogenic cell death and increase in effector memory T cells by the MIMIC combination therapy . (A) Representative immunofluorescent staining of calreticulin (green) and E-cadherin (red) in subcutaneous and peritoneal tumour tissues of treatment groups receiving Ox, Ox + ICI + Luc-mRNA, and Ox + ICI + IFNα/IL12-mRNA in the FC1245 model. Ox: oxaliplatin. ICI: immune checkpoint inhibitors consisting of anti-PD-1 antibody and anti-CTLA-4 antibody. (B, C) Membrane-to-cytoplasm ratio of calreticulin (CRT) fluorescence intensity in subcutaneous (B) and peritoneal (C) tumour tissues. P-values were calculated by ANOVA with the Tukey–Kramer post hoc test. (D) HMGB1 level in peritoneal lavage fluid. P-values were calculated by ANOVA with the Tukey–Kramer post hoc test. (E, F) Representative flow cytometry plots (E) and quantification (F) of CD8+ CD44+ CD62L-effector memory T (Tem) cells in peripheral blood. P-values were calculated by ANOVA with the Tukey–Kramer post hoc test. ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001.

Article Snippet: The HMGB1 concentration in the lavage fluid was quantified using HMGB1 ELISA kit (Arigo Biolaboratories, Hsinchu, Taiwan), and the total amount of HMGB1 in the peritoneal cavity was calculated based on the measured concentration and the lavage volume.

Techniques: Staining, Membrane, Fluorescence, Flow Cytometry