Journal: Journal of Nanobiotechnology
Article Title: Multifunctional nanoagonist enhances photodynamic therapy-driven in situ cancer vaccination by inhibiting tumor thrombosis
doi: 10.1186/s12951-025-03843-8
Figure Lengend Snippet: In vitro evaluation of cDVPMA-induced ICD effects. ( A ) Representative CLSM images of CRT exposure on 4T1 cells following different treatments. Scale bar = 20 μm. ( B ) Representative CLSM images of HMGB1 release from 4T1 cells following different treatments. Scale bar = 20 μm. ( C ) Quantitative analysis of CRT exposure on 4T1 cells following different treatments. ( D ) Quantitative analysis of HMGB1 release from 4T1 cells following different treatments. ELISA analysis of ( E ) HMGB1 and ( F ) ATP release from 4T1 cells following different treatments. ( G ) Western blot analysis and quantification of ( H ) p-STING activation, ( I ) p-TBK1 activation, ( J ) p-IRF3 activation after various treatments. ( K ) Scheme illustration of the transwell co-culture system. Concentrations of ( L ) IFN-β and ( M ) TNF-α in the supernatants of different treatment groups. ( N ) Quantitative analysis and ( O ) representative flow cytometry plots of the proportion of mature DCs after co-incubation with 4T1 cell supernatants pre-stimulated with different formulations. FL., fluorescence intensity. c, free 2′3′-cGAMP. All data are represented as mean ± SD ( n = 3). ns, no significance. * p < 0.05, ** p < 0.01, *** p < 0.001
Article Snippet: Singlet Oxygen Sensor Green (SOSG) fluorescent probe, dichlorodihydrofluorescein diacetate (DCFH-DA), cytokine detection kits (IFN-β, IFN-γ, IL-6, TNF-α, IL-1β, TGF-β1), ATP detection kit, 4′,6-diamidino-2-phenylindole (DAPI), protease inhibitors, Alexa Fluor 555-labeled donkey anti-rabbit IgG (H + L), rabbit anti-mouse β -actin monoclonal antibody, HRP-labeled goat anti-rabbit IgG (H + L), HMGB1 detection kit, GM-CSF, and IL-4 were all purchased from Shanghai Bio-Tech Biotechnology Co., Ltd. Rabbit anti-mouse calreticulin polyclonal antibody and rabbit anti-mouse HMGB1 polyclonal antibody were purchased from Proteintech Group, Inc. Fetal bovine serum (FBS), Dulbecco’s modified Eagle’s medium (DMEM), penicillin, streptomycin, and trypsin were obtained from Thermo Fisher Scientific Inc. Phospho-STING (Ser365) (D8F4W) rabbit monoclonal antibody (#72971), and Phospho-IRF-3 (Ser396) (4D4G) rabbit monoclonal antibody (#4947) were purchased from Cell Signaling Technology, Inc. Anti-NAK/TBK1 (phospho S172) antibody [EPR2867(2)] was obtained from Abcam.
Techniques: In Vitro, Enzyme-linked Immunosorbent Assay, Western Blot, Activation Assay, Co-Culture Assay, Flow Cytometry, Incubation, Fluorescence
