Journal: iScience

Article Title: Direct interaction of HMGB1 with SARS-CoV-2 facilitates its infection via RAGE-dependent endocytosis

doi: 10.1016/j.isci.2025.113063

Figure Lengend Snippet: HMGB1 induces SARS-CoV-2 infection in an ACE2-independent RAGE-dependent manner (A) Western blot analysis of receptors responsible for SARS-CoV-2 infection and HMGB1 binding. S.E., short exposure; L.E., long exposure. (B) Flow cytometry analysis of ectodomain ACE2 in A549 and NCI-H1975 cells used in this study. Vero E6 cells were used as control. (C) A549 cells were transfected with shRNA-ACE2 for 48 h prior to infection with 1 MOI SARS-CoV-2, which was preincubated with HMGB1 for 1 h at 37°C, cultured further for 3 h, and subjected to western blotting. (D and E) A549 cells were infected for 1 h with 1 MOI SARS-CoV-2, which was preincubated with HMGB1 for 1 h, in the presence of 40 μg/mL sRAGE at 37°C as indicated. Cells were harvested at 3 hpi and subjected to western blotting (D) n = 3, and qRT-PCR for viral RNA measurement (E) n = 3. (F) NCI-H1975 cells were infected with SARS-CoV-2 preincubated with 20 μg/mL HMGB1 in the presence of azeliragon. Cells were harvested at 3 hpi and subjected to western blotting. (G) A549 cells were infected with 5 MOI SARS-CoV-2, which was treated as above to observe NP. Representative confocal images are shown. The percentage of infected cells and NP intensity were measured by counting at least 700 visible cells. n = 4. (H) Cycloheximide pretreated NCI-H1975 cells were infected with 5 MOI SARS-CoV-2 (preincubated with HMGB1). Cells were stained for NP before permeabilization (green; external) and post-permeabilization (red; external and internal). Representative images and their magnifications are shown. The percentage of intracellular spots was measured by counting at least 200 visible cells. n = 4. (I and J) SARS2pp was preincubated with or without HMGB1 in the presence of sRAGE before transduction in NCI-H1975 cells. NanoLuc luciferase activity was measured 72 h post-transduction. n = 3 Scale bars represent 5 μm. Data are presented as mean ± SEM, ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001; ns, not significant, using one-way ANOVA with Tukey’s multiple comparison test and Student’s unpaired t-test. HMGB1, high-mobility group box 1; SARS-CoV-2, severe acute respiratory syndrome coronavirus 2; MOI, multiplicity of infection; qRT-PCR, quantitative reverse transcription polymerase chain reaction; NP, nucleocapsid protein; SEM, standard error of the mean; ANOVA, analysis of variance; hpi, hours post-infection; RAGE, receptor for advanced glycation end-products; ACE2, angiotensin-converting enzyme 2; SARS2pp, SARS-CoV-2 spike protein (S)-pseudotyped retrovirus.

Article Snippet: 40 μg/mL sRAGE (11629-HCCH, Sino Biological, Oklahoma City, OK, USA), azeliragon (S6415, Selleckchem, Houston, TX, USA), dynasore (D7693, Sigma-Aldrich, St. Louis, MO, USA) and chloroquine (C6628, Sigma-Aldrich) were pretreated for 2 h at 37°C, prior to the infection procedure.

Techniques: Infection, Western Blot, Binding Assay, Flow Cytometry, Control, Transfection, shRNA, Cell Culture, Quantitative RT-PCR, Staining, Transduction, Luciferase, Activity Assay, Comparison, Reverse Transcription, Polymerase Chain Reaction

Journal: BMJ Open Respiratory Research

Article Title: Validation of plasma soluble receptor of advanced glycation end-products and angiopoietin-2 in paediatric acute respiratory distress syndrome

doi: 10.1136/bmjresp-2025-003630

Figure Lengend Snippet: Plasma sRAGE and Ang-2 levels stratified by PARDS severity Comparison was made between mild, moderate and severe PARDS versus control using the Mann-Whitney U test. P values are subjected to Bonferroni correction for multiple testing. Ang-2, angiopoietin-2; PARDS, paediatric acute respiratory distress syndrome; sRAGE, soluble receptor for advanced glycation end-products.

Article Snippet: The plasma was then aspirated and stored at −80°C for later batched analysis. sRAGE (R&D Systems; catalogue no. DY1145) and Ang-2 (R&D Systems; catalogue no. DY623) were measured in duplicate using an ELISA according to manufacturer’s protocol.

Techniques: Clinical Proteomics, Comparison, Control, MANN-WHITNEY

Journal: BMJ Open Respiratory Research

Article Title: Validation of plasma soluble receptor of advanced glycation end-products and angiopoietin-2 in paediatric acute respiratory distress syndrome

doi: 10.1136/bmjresp-2025-003630

Figure Lengend Snippet: Correlation matrix of plasma sRAGE and Ang-2 with age, PIM 3 score, PELOD2 score and the oxygenation index. Spearman test was used to assess correlation. Numbers in cells refer to Spearman’s correlation coefficient. *p<0.05, **p<0.01, ***p<0.001. Ang-2, angiopoietin-2; PELOD 2, Pediatric Logistic Organ Dysfunction 2 score; PIM 3, Pediatric Index of Mortality 3 score; sRAGE, soluble receptor for advanced glycation end-products.

Article Snippet: The plasma was then aspirated and stored at −80°C for later batched analysis. sRAGE (R&D Systems; catalogue no. DY1145) and Ang-2 (R&D Systems; catalogue no. DY623) were measured in duplicate using an ELISA according to manufacturer’s protocol.

Techniques: Clinical Proteomics

Journal: BMJ Open Respiratory Research

Article Title: Validation of plasma soluble receptor of advanced glycation end-products and angiopoietin-2 in paediatric acute respiratory distress syndrome

doi: 10.1136/bmjresp-2025-003630

Figure Lengend Snippet: Plasma sRAGE and Ang-2 levels in the presence of cardiovascular dysfunction, multiorgan dysfunction and mortality. Data presented as median (IQR), and comparison was made using the Mann-Whitney test. Organ dysfunctions were defined according to the International Pediatric Sepsis Consensus Conference. Ang-2, angiopoietin-2; ICU, intensive care unit; sRAGE, soluble receptor for advanced glycation end-products.

Article Snippet: The plasma was then aspirated and stored at −80°C for later batched analysis. sRAGE (R&D Systems; catalogue no. DY1145) and Ang-2 (R&D Systems; catalogue no. DY623) were measured in duplicate using an ELISA according to manufacturer’s protocol.

Techniques: Clinical Proteomics, Comparison, MANN-WHITNEY

Journal: Physiological Reports

Article Title: Biological sex modulates lung injury severity in adolescent mice exposed to short‐term aerosolized vitamin E acetate

doi: 10.14814/phy2.70691

Figure Lengend Snippet: Serum sRAGE concentration (ng/mL) in the BALF supernatant was measured using ELISA. 3‐day and 10‐day VEA‐exposed males and females displayed significantly higher serum sRAGE levels compared to control groups. Additionally, 3‐day and 10‐day VEA‐exposed females presented with significantly higher sRAGE levels when compared to their male counterparts. Column height represents mean concentration ± standard error (SE). N = 6/treatment group. BALF, bronchoalveolar lavage fluid; sRAGE, soluble receptor of advanced glycation end products; VEA, vitamin E acetate.

Article Snippet: Serum sRAGE levels were quantified using the murine sRAGE Quantikine ELISA kit (Cat. NO. MRG00, R&D Systems, Minneapolis, MN, USA) following the manufacturer's instructions.

Techniques: Concentration Assay, Enzyme-linked Immunosorbent Assay, Control

Journal: medRxiv

Article Title: Aerobic Exercise Training Elevates Circulating sRAGE via Modulation of Sheddase Regulation in Adults with Type 2 Diabetes

doi: 10.1101/2025.09.10.25335517

Figure Lengend Snippet: Basal solubilized receptor for advanced glycation end products (sRAGE; A), solubilized Toll-like receptor 4 (sTLR4, C) pre and post 12-week intervention in the control (CON) and aerobic exercise training (AE) groups. Respective absolute changes in basal sRAGE (B) and sTLR4 (C) and percentage changes in sRAGE (E) and sTLR4 (E) were calculated. *P < 0.05 vs Pre, # P < 0.05 vs CON. Statistical analyses performed include mixed model regression analyses with Bonferroni post hoc (panels A & C) and two-tailed t-test (panels B, D, & E).

Article Snippet: sRAGE was quantified via sRAGE ELISA (DRG000, R&D Systems, Minneapolis, MN) according to manufacturer’s instructions.

Techniques: Control, Two Tailed Test