Journal: Journal of Cellular and Molecular Medicine

Article Title: Exosomal HMGB1 Orchestrates NSCLC Progression and Immunosuppressive Macrophage Polarisation Through the TLR4 / NF ‐ κB / IL ‐6/ STAT3 Signalling Cascade

doi: 10.1111/jcmm.71050

Figure Lengend Snippet: Exosomal HMGB1 activates JAK/STAT3 signalling to promote NSCLC progression. (A) Protein–protein interaction (PPI) network analysis of HMGB1 using the STRING database. (B) Western blot analysis of NF‐κB in A549 and PC9 cells treated with PBS, recombinant HMGB1 (100 ng), exosomes from vector cells or exosomes from HMGB1 OE cells (cell‐to‐exosome ratio = 1:10). (C) ELISA quantification of IL‐6 in the supernatant of A549 and PC9 cells under the same treatment conditions as in (B). (D) Immunofluorescence staining of p‐STAT3 of A549 and PC9 cells under the same treatments, including an additional group co‐treated with exosomes from HMGB1 OE cells and NF‐κB inhibitor (50 μM). (E) Cell proliferation of A549 and PC9 cells treated with HMGB1 OE‐derived exosomes alone or in combination with NF‐κB inhibitor (50 μM) or STAT3 inhibitor (20 μM). (F) Cell migration under the same treatment conditions as in (E). (G) Colony formation assays of A549 and PC9 cells under the same treatment conditions as in (E).

Article Snippet: IL‐6 neutralising antibodies were obtained from Sino Biological (China).

Techniques: Western Blot, Recombinant, Plasmid Preparation, Enzyme-linked Immunosorbent Assay, Immunofluorescence, Staining, Derivative Assay, Migration

Journal: Journal of Cellular and Molecular Medicine

Article Title: Exosomal HMGB1 Orchestrates NSCLC Progression and Immunosuppressive Macrophage Polarisation Through the TLR4 / NF ‐ κB / IL ‐6/ STAT3 Signalling Cascade

doi: 10.1111/jcmm.71050

Figure Lengend Snippet: Targeting HMGB1 signalling improves therapeutic outcomes in NSCLC. (A) Correlation analysis between immune infiltration scores and HMGB1 expression in 491 LUAD and 500 LUSC patients from the TCGA database. (B) Correlation between HMGB1 expression and the distribution of various immune cell subsets in LUAD and LUSC patients. (C, D) THP‐1–derived M0 macrophages were treated with PBS, HMGB1 (10 or 100 ng) or exosomes derived from vector or HMGB1 OE cells (cell‐to‐exosome ratio = 1:10). M1 macrophage markers (CD86, CD80, iNOS) and M2 markers (CD206, IL‐10, Arg1) were quantified by PCR. (E) Lewis tumour‐bearing mice were treated with PBS, HMGB1 OE‐derived exosomes (1 × 10 10 exosomes per mouse, twice per week), anti‐PD‐1 antibody (RMP1‐14, 200 μg per mouse, twice per week) or combination therapy ( n = 5 per group). Tumour volumes and apoptosis levels in tumour tissues (day 25) were assessed. (F) PC9 cells were treated with PBS or exosomes from HMGB1 OE cells (cell‐to‐exosome ratio = 1:10), followed by Osimertinib (50 nM, 48 h), and apoptosis was measured. (G) A549 and PC9 cells were similarly treated with PBS or HMGB1 OE‐derived exosomes, followed by Cisplatin (5 μM, 48 h), and apoptosis was analysed. (H) A549 and PC9 cells were similarly treated with paclitaxel (10 μM, 48 h) under the same conditions, and cell apoptosis was determined. (I) A549‐bearing mice were treated with HMGB1 OE‐derived exosomes (1 × 10 10 exosomes per mouse), followed by PBS, paclitaxel (PTX, 10 mg/kg, twice per week), STAT3 inhibitor (5 mg/kg, twice per week) or combination therapy. (J) Schematic diagram illustrating the proposed mechanism: HMGB1 upregulates TLR4, thereby activating the NF‐κB–IL‐6 axis and stimulating JAK2/STAT3 signalling to promote tumour progression. Concurrently, HMGB1 facilitates M2 macrophage polarisation.

Article Snippet: IL‐6 neutralising antibodies were obtained from Sino Biological (China).

Techniques: Expressing, Derivative Assay, Plasmid Preparation

Journal: Redox Biology

Article Title: The electrophilic metabolite of kynurenine, kynurenine-CKA, requires C151 in Keap1 to derepress Nrf2

doi: 10.1016/j.redox.2026.104009

Figure Lengend Snippet: AhR inhibition has no effect on the anti-inflammatory actions of Kyn-CKA in BMDMs. (A) Mouse AhR reporter cells expressing luciferase under the control of the xenobiotic response element (XRE) were incubated with Kyn-CKA or kynurenine for 24 h prior to luminescence measurement (n = 3). Data were fitted to a sigmoidal four-parameter logistic curve (L-Kyn: r 2 = 0.824, IC 50 = 28 μM, span = 3218 RFU. Kyn-CKA: r 2 = 0.970, IC 50 = 13 μM, span = 47,901 RFU). (B) Inhibition of AhR-dependent luciferase expression by CH223191 (AhRinh). (C–F) Kyn-CKA inhibits the expression of the NF-κB-regulated genes IL6, MCP1, Nos2 and IL1β in BMDM following 5 h incubation with LPS (0.5 μg/mL) and the indicated concentrations of Kyn-CKA both in the presence or absence of CH223191 (10 μM). Data are n = 3–4, ∗∗∗∗p < 0.0001 by one-way ANOVA and Tukey's post-test. (G – H) Extracellular cytokine levels from BMDM treated as in C–F for 5 h. Data are n = 3–4 (all replicates shown), ∗∗∗∗p < 0.0001 by one-way ANOVA and Tukey's post-test.

Article Snippet: The TaqManTM Gene Expression Assay IDs (Thermo Fisher, Waltham, MA) were: Mm01253561 (for Nqo1 ); Mm00516005 (for Ho1); Mm01324400 (for Gclm ); Mm00434228 (for IL1b ); Mm00446190 (for IL6 ); Mm00441242 (for Mcp1 ); Mm00443252 (for Tnf ); Mm004405021 (for Nos2 ); Mm00437762 (for B2m ); Mm03024075 (for Hprt1 ); and 4352341E (for Actb ).

Techniques: Inhibition, Expressing, Luciferase, Control, Incubation

Journal: Redox Biology

Article Title: The electrophilic metabolite of kynurenine, kynurenine-CKA, requires C151 in Keap1 to derepress Nrf2

doi: 10.1016/j.redox.2026.104009

Figure Lengend Snippet: The low-dose anti-inflammatory effects of Kyn-CKA in BMDMs are dependent on Nrf2. (A – B) Treatment with Kyn-CKA (5 h) fails to induce the expression of Nqo1 and Gclm in BMDMs obtained from Nrf2-knockout (Nrf2 −/− ) mice. Data are n = 3, ∗∗∗∗p < 0.0001 by two-way ANOVA and Tukey's post-test. (C – G) Effects of Kyn-CKA on the expression of NF-κB−regulated genes in WT and Nrf2-KO BMDMs following 5 h incubation with LPS (0.5 μg/mL) and the indicated concentrations of Kyn-CKA. Data are n = 6, ∗p < 0.05, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001 by two-way ANOVA and Tukey's post-test. Nrf2 was identified as a significant source of variation for MCP1, IL6, TNFα, Nos2, (p < 0.0001) and IL1β (p < 0.005).

Article Snippet: The TaqManTM Gene Expression Assay IDs (Thermo Fisher, Waltham, MA) were: Mm01253561 (for Nqo1 ); Mm00516005 (for Ho1); Mm01324400 (for Gclm ); Mm00434228 (for IL1b ); Mm00446190 (for IL6 ); Mm00441242 (for Mcp1 ); Mm00443252 (for Tnf ); Mm004405021 (for Nos2 ); Mm00437762 (for B2m ); Mm03024075 (for Hprt1 ); and 4352341E (for Actb ).

Techniques: Expressing, Knock-Out, Incubation