Journal: Frontiers in Public Health

Article Title: Proton beam therapy induces protective immunity via HMGB1-dependent signaling

doi: 10.3389/fpubh.2025.1686678

Figure Lengend Snippet: The expression of proton-induced CRT and HMGB1. (A,C,D) Flow cytometry analysis of CRT at 24 h post-proton irradiation. (B,E,F) Flow cytometry analysis of CRT at 48 h post-proton irradiation. (G) The temporal characteristics of proton-induced CRT expression. (H–J) The expression of HMGB1 in the supernatant of cell culture medium 24 (I) and 48 (J) hours after proton irradiation. (H) The temporal characteristics of proton-induced HMGB1 expression. MFI means median fluorescence intensity. * means p < 0.05, ** means p < 0.01,*** means p < 0.001, ns means not significant.

Article Snippet: HMGB1 and Interferon-gamma (IFN-γ) in the mouse serum samples were measured by using HMGB1 kit (H257-1-2, Mouse HMGB1 ELISA Kit, NJJC Bioscience, Nanjing, China) and IFN-γ kit (#EK0375, Boster Biological Technology, Wuhan, China) according to the manufacturer’s instructions.

Techniques: Expressing, Flow Cytometry, Irradiation, Cell Culture, Fluorescence

Journal: Frontiers in Public Health

Article Title: Proton beam therapy induces protective immunity via HMGB1-dependent signaling

doi: 10.3389/fpubh.2025.1686678

Figure Lengend Snippet: Proton radiation stimulates immunogenic cell death. (A) Experimental schedule for proton-induced ICD evaluation. Primary tumor growth (B) and distal tumor growth (C) after the transplantation of different doses (3, 6, 12 Gy) proton-irradiated cells. Four weeks after tumor cell transplantation, spleens were harvested and analysed: immunohistochemical staining of T lymphocytes (D) and IFN- γ (G) infiltration in the spleen, and quantification by flow cytometry (E,F) . (H) The expression of HMGB1 in serum after the transplantation of proton-irradiated cells. * means p < 0.05, ** means p < 0.01,*** means p < 0.001, **** means p < 0.0001, ns means not significant.

Article Snippet: HMGB1 and Interferon-gamma (IFN-γ) in the mouse serum samples were measured by using HMGB1 kit (H257-1-2, Mouse HMGB1 ELISA Kit, NJJC Bioscience, Nanjing, China) and IFN-γ kit (#EK0375, Boster Biological Technology, Wuhan, China) according to the manufacturer’s instructions.

Techniques: Transplantation Assay, Irradiation, Immunohistochemical staining, Staining, Flow Cytometry, Expressing

Journal: Frontiers in Public Health

Article Title: Proton beam therapy induces protective immunity via HMGB1-dependent signaling

doi: 10.3389/fpubh.2025.1686678

Figure Lengend Snippet: Verification of silencing of shCRT and shHMGB1 cell lines by proton irradiation. The mRNA expression level of CRT (A) and HMGB1 (B) after corresponding gene silencing. Surface expression of CRT (C) and secreted HMGB1 concentration (D) in CRT- or HMGB1-knockdown Colon-26 stable cell lines were measured after irradiation by flow cytometry and ELISA, respectively. * means p < 0.05, ** means p < 0.01,*** means p < 0.001, **** means p < 0.0001, ns means not significant.

Article Snippet: HMGB1 and Interferon-gamma (IFN-γ) in the mouse serum samples were measured by using HMGB1 kit (H257-1-2, Mouse HMGB1 ELISA Kit, NJJC Bioscience, Nanjing, China) and IFN-γ kit (#EK0375, Boster Biological Technology, Wuhan, China) according to the manufacturer’s instructions.

Techniques: Irradiation, Expressing, Concentration Assay, Knockdown, Stable Transfection, Flow Cytometry, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Public Health

Article Title: Proton beam therapy induces protective immunity via HMGB1-dependent signaling

doi: 10.3389/fpubh.2025.1686678

Figure Lengend Snippet: The role of Proton-induced CRT and HMGB1 in distal tumor colonization rejection. (A) Experimental schedule for CRT and HMGB1 evaluation. (B) Tumor growth after the transplantation of 12 Gy Proton-irradiated cells. The black dotted line indicates mean volume of non-irradiated treated tumor. The blue solid line indicates that the volume of the distal tumor is close to zero, meaning a state of complete immunity. The orange solid line indicates that the volume of the distal tumor is above zero yet remains below those of untreated controls, meaning a state of partial immune control. (C) The expression of HMGB1 in serum after the transplantation of 12 Gy Proton-irradiated cells. Ratio of CD4 positive T cell.

Article Snippet: HMGB1 and Interferon-gamma (IFN-γ) in the mouse serum samples were measured by using HMGB1 kit (H257-1-2, Mouse HMGB1 ELISA Kit, NJJC Bioscience, Nanjing, China) and IFN-γ kit (#EK0375, Boster Biological Technology, Wuhan, China) according to the manufacturer’s instructions.

Techniques: Transplantation Assay, Irradiation, Control, Expressing

Journal: Advanced Science

Article Title: Engineered Bacteria‐Vesicle Delivered Lactate Reprogramming Boosts Tumor Radiosensitivity

doi: 10.1002/advs.202524303

Figure Lengend Snippet: In vitro cytotoxicity and mechanistic analysis of LOx‐loaded OMVs. (a) TEM characterization of OMVs‐P (OMVs derived from ENHP) and OMVs‐L (OMVs derived from ENHL) morphology; scale bar: 200 nm. (b,c) Flow cytometric analysis of CT26 cell apoptosis: representative plots (b) and quantitative analysis (c) following treatments (G1: PBS control; G2: OMVs‐P; G3: OMVs‐L; G4: 6 Gy; G5: OMVs‐P + 6 Gy; G6: OMVs‐L + 6 Gy; n = 5). (d) Cell viability assessment of CT26 cells post‐treatment ( n = 5). (e) Quantification of colony counts with different treatments towards CT26 cells. (f) Immunofluorescence images of γ‐H2AX staining after different treatments; scale bar: 100 µm. (g) The lactate content in the supernatant of CT26 cells after different treatments ( n = 5). (h) Western blot analysis of CT26 cell lysates for Pan‐Kla, GAPDH, and MRE11 expression, along with Pan‐Kla‐modified MRE11 levels under different treatments. All data represent mean ± SD. Statistical significance was determined by one‐way ANOVA with Tukey's multiple‐comparisons test (* p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, ns. not significant).

Article Snippet: Primary antibodies included anti‐Syndecan‐1 antibody (Abcam, ab128936), anti‐Dnak antibody (Abcam, ab69617), Anti‐L‐Lactyl Lysine Rabbit mAb (PTM Bio, 9H1L6), Ki‐67 Polyclonal antibody (Proteintech, 28074‐1‐AP), HIF1α Rabbit pAb (ABclonal, A11945), LDHA Rabbit pAb (ABclonal, A1146), β‐Actin Rabbit mAb (ABclonal, AC050), DDDDK‐Tag Rabbit mAb (ABclonal, AE092), Myc‐Tag Rabbit mAb (ABclonal, AE070), GAPDH Rabbit mAb (ABclonal, A19056), MRE11 (Boster, BM5057), and Anti‐gamma H2A.X (Abcam, ab81299).

Techniques: In Vitro, Derivative Assay, Control, Immunofluorescence, Staining, Western Blot, Expressing, Modification