phospho irf3  (Affinity Biosciences)

Journal: Bioactive Materials

Article Title: On-demand mild photothermal cascade platform reprogramming mitochondrial immunity for tendon rejuvenation

doi: 10.1016/j.bioactmat.2026.01.004

Figure Lengend Snippet: Schematic illustration of the ROS-responsive on-demand mild photothermal cascade platform for tendon rejuvenation in Achilles tendinopathy. The platform strategically modulates the mitochondrial-cGAS-STING-IRF3/NF-κB signaling axis and induces heat shock protein 70 (HSP70) expression to attenuate oxidative stress and cellular senescence within tendon stem/progenitor cells (TSPCs). Consequently, it promotes tenogenic differentiation while abrogating aberrant osteogenic/chondrogenic lineage commitment. This cascade effect ultimately mitigates heterotopic ossification, enhances structural tendon regeneration, restores biomechanical function, and alleviates pain. Nanoparticle nomenclature: LA-NPs and TPA-TCNQ-NPs denote nanoparticles encapsulating LA or TPA-TCNQ individually; LT-NPs refers to the composite formulation consisting of a mixture of LA-NPs and TPA-TCNQ-NPs; and LT-NPs-NIR represents the LT-NPs mixture following 808 nm near-infrared (NIR) irradiation to activate the photothermal response and controlled payload release.

Article Snippet: After blocking for 1 h, membranes were incubated overnight at 4 °C with primary antibodies against STING (13647, CST, USA; A21051, Abclonal, China), p-STING (72971, CST, USA; AF7416, Affinity, China), IRF3 (ab68481, Abcam, UK), p-IRF3 (29047, CST, USA), P65 (A22331, Abclonal, China; 8242, CST, USA), p-P65 (AP0124, Abclonal, China), P53 (10442-1-AP, Proteintech, USA), SOX9 (sc-166505, Santa Cruz, USA), BMP-2 (ab284387, abcam, USA), OCN (sc-390877, Santa Cruz, USA), and iNOS (ab178945, Abcam, USA).

Techniques: Expressing, Formulation, Irradiation

Journal: Bioactive Materials

Article Title: On-demand mild photothermal cascade platform reprogramming mitochondrial immunity for tendon rejuvenation

doi: 10.1016/j.bioactmat.2026.01.004

Figure Lengend Snippet: LT-NPs-NIR regulate the mtDNA-STING-IRF3/NF-κB pathway in macrophages. (A) Principal component analysis (PCA) of transcriptomic data from RAW 264.7 cells under different treatments. (B) Volcano plots comparing H 2 O 2 vs. Control (left) and LT-NPs-NIR vs. H 2 O 2 (right). (C) Heatmap of differentially expressed genes (DEGs) with hierarchical clustering. (D, E) Gene Ontology (GO), KEGG, and Gene Set Enrichment Analysis (GSEA) for H 2 O 2 vs. Control (D) and LT-NPs-NIR vs. H 2 O 2 (E). (F) Multi-SIM imaging showing mtDNA (magenta) and the mitochondrial outer membrane protein TOMM20 (green), with quantification of their colocalization. (G) Western blot analysis of key proteins in the cGAS-STING-NF-κB axis. Scale bar: 5 μm (F). Significance: ns, not significant; ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001.

Article Snippet: After blocking for 1 h, membranes were incubated overnight at 4 °C with primary antibodies against STING (13647, CST, USA; A21051, Abclonal, China), p-STING (72971, CST, USA; AF7416, Affinity, China), IRF3 (ab68481, Abcam, UK), p-IRF3 (29047, CST, USA), P65 (A22331, Abclonal, China; 8242, CST, USA), p-P65 (AP0124, Abclonal, China), P53 (10442-1-AP, Proteintech, USA), SOX9 (sc-166505, Santa Cruz, USA), BMP-2 (ab284387, abcam, USA), OCN (sc-390877, Santa Cruz, USA), and iNOS (ab178945, Abcam, USA).

Techniques: Control, Imaging, Membrane, Western Blot

Journal: Bioactive Materials

Article Title: On-demand mild photothermal cascade platform reprogramming mitochondrial immunity for tendon rejuvenation

doi: 10.1016/j.bioactmat.2026.01.004

Figure Lengend Snippet: LT-NPs-NIR attenuate oxidative stress, preserve mitochondrial integrity, and suppress senescence in TSPCs by inhibiting the mtDNA-STING-NF-κB axis. (A) Schematic of the experimental design. (B) TSPC proliferation assessed by CCK-8 assay. (C, E) Immunofluorescence staining and quantification of HSP70 (n = 3). (D, F) Mitochondrial membrane potential (ΔΨm) visualized by JC-1 staining (red: high potential; green: low potential) and quantification. (G) Multi-SIM of mtDNA (magenta) and TOMM20 (green) with colocalization analysis. (H) Western blot of cGAS-STING-IRF3-NF-κB pathway proteins. (I, K) Colony-forming unit fibroblast (CFU-F) assay and quantification of self-renewal capacity (n = 3). (J) Senescence-associated β-galactosidase (SA-β-gal) activity. (L, M) Apoptosis analysis by Annexin V/PI flow cytometry and quantification (n = 3). Scale bars: 100 μm (C); 5 μm (D, G); 200 μm (J). Significance: ns, not significant; ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001.

Article Snippet: After blocking for 1 h, membranes were incubated overnight at 4 °C with primary antibodies against STING (13647, CST, USA; A21051, Abclonal, China), p-STING (72971, CST, USA; AF7416, Affinity, China), IRF3 (ab68481, Abcam, UK), p-IRF3 (29047, CST, USA), P65 (A22331, Abclonal, China; 8242, CST, USA), p-P65 (AP0124, Abclonal, China), P53 (10442-1-AP, Proteintech, USA), SOX9 (sc-166505, Santa Cruz, USA), BMP-2 (ab284387, abcam, USA), OCN (sc-390877, Santa Cruz, USA), and iNOS (ab178945, Abcam, USA).

Techniques: CCK-8 Assay, Immunofluorescence, Staining, Membrane, Western Blot, Activity Assay, Flow Cytometry