mouse rabbit ig  (Vector Laboratories)

Journal: Regenerative Therapy

Article Title: Inhibition of the TLR4/RAGE pathway by clearance of extracellular HMGB1 is a potential therapeutic target for radiation-damaged salivary glands

doi: 10.1016/j.reth.2025.07.004

Figure Lengend Snippet: Histological observations of TLR4 and RAGE expression in SGs at 12 weeks post-IR. (A) Expression of TLR4 (red, TLR4; blue, DAPI; scale bar, 100 μm) ( × 200). (B) Expression of TLR4 and AQP5 (red, TLR4; green, AQP5; blue, DAPI; scale bar, 100 μm) (upper, × 200; lower, × 400). (C) Expression of TLR4 and KRT14 (red, TLR4; green, KRT14; blue, DAPI; scale bar, 100 μm) (upper, × 200; lower, × 400). (D) Expression of RAGE (red, RAGE; blue, DAPI; scale bar, 100 μm) (upper, × 200; lower, × 400). (E) Expression of RAGE and CD31 (red, CD31; green, RAGE; blue, DAPI; scale bar, 20 μm) ( × 1000). (F) Expression of RAGE and F4/80 (red, RAGE; green, F4/80; blue, DAPI; scale bar, 20 μm) ( × 1000).

Article Snippet: In brief, immunofluorescence analyses were performed using rabbit anti-mouse TLR4 antibody (1:100; abcam, MA, USA), mouse anti-mouse Aquaporin-5 (AQP5) antibody (1:100; Santa Cruz Biotechnology, CA, USA), rabbit anti-mouse Cytokeratin-14 (KRT14) antibody (1:100; Proteintech, IL, USA), rabbit anti-mouse TLR2 antibody (1:100; abcam), rabbit anti-mouse RAGE antibody (1:50; abcam), rabbit anti-mouse CD31 antibody (1:100; abcam), rabbit anti-mouse F4/80 antibody (1:50; abcam), and mouse anti-mouse phospho-nuclear factor-kappa B (p–NF–κB) antibody (1:50; Santa Cruz Biotechnology).

Techniques: Expressing

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