Fig 2: Increased DNA damage and ER stress in cells treated with NP-2 under NIR II light irradiation. Expression of PARP, XBP1, HMGB1 protein in cells treated with Cisplatin, NP-1, NP-2, NP-2+L (A). Quantification of PARP, XBP1, and HMGB1 level in A (B). Quantification of the XBP1 level in A2780 cells via flow cytometry (C). CLSM images showing translocation of XBP1 in A2780 cells treated with Cisplatin, NP-1, NP-2, and NP-2+L respectively (D). Significance is defined as ****p < 0.0001.

Fig 3: The serum level of secreted HMGB1 predicts response to treatment with bevacizumab(A) 20 established renal cancer PDX models were characterized bytreatment with anti-VEGF monoclonal antibody bevacizumab. Tumor volume was determined twice weekly until animals of the control group reached termination criteria. Group median absolute tumor volumes are plotted over time. Tumors with high secretion of HMGB1 (upper panel) responded significantly better to treatment with bevacizumab than tumors with low secretion of HMGB1 (lower panel). (B) The optimal T/C values were plotted as waterfall plot for 20 renal cancer PDX models treated with bevacizumab. The blue bars represent HMGB1 low secretor models; the green bars represent HMGB1 high secretor models. The optimal T/C values of the high secretor group were significantly lower (p < 0.0009, Mann–Whitney test, two-tailed) as the respective values of the low secretor group. (C) The optimal T/C values were plotted as waterfall plot for 20 renal cancer PDX models treated with bevacizumab. The purple bars represent human VEGFA low expressing models; the yellow bars represent human VEGFA high expressing models. The difference between the two groups was statically significant (p< 0.05, Mann–Whitney test, two-tailed). Thus, the level of secreted HMGB1 is a stronger predictive marker for bevacizumab sensitivity as the expression of human VEGFA.

Fig 4: HMGB1 triggered ERK-mediated Drp1 phosphorylation via RAGE in CRC.a SW480 and LoVo cells were treated with the recombinant human HMGB1 protein (0, 1, and 2 µg/mL) for 24 h. Cell lysate was analyzed by immunoblotting. Quantification of these results is shown (n = 3). ***p < 0.001. b SW480 cells were treated with the recombinant human HMGB1 protein (1 µg/mL) and RAGE inhibitor (10 µM FPS-ZM1) for 24 h. Cell lysate was analyzed by immunoblotting. Quantification of these results is shown (n = 3). **p < 0.01 and ***p < 0.001. c SW480 cells were transfected with the pcDNA3.1-vector, pcDNA3.1-wtRAGE for 24 h and then treated with the recombinant human HMGB1 protein (1 µg/mL) for 24 h. Cell lysate was analyzed by immunoblotting. Quantification of these results is shown (n = 3). **p < 0.01. d SW480 cells were silenced with lentivirus carrying shRNA again RAGE, and then treated with the recombinant human HMGB1 protein (1 µg/mL) for 24 h. Cell lysate was analyzed by immunoblotting. e LoVo cells were treated with recombinant human HMGB1 (1 µg/mL) for 24 h. Cell lysate was analyzed by immunoblotting. Quantification of these results is shown (n = 3). ***p < 0.001. Mitochondria morphology was observed by MitoTracker Red staining. Scale bar = 10 µm. f LoVo cells were treated with RAGE inhibitor (10 µM FPS-ZM1), Drp1 inhibitor (50 µM Mdivi-1) and recombinant human HMGB1 (1 µg/mL) for 1 h prior to addition of different dosages of OXP for 48 h. The cell viability was analyzed using an MTT assay (n = 3). *p < 0.05. g LoVo cells were treated with RAGE inhibitor (10 µM FPS-ZM1), Drp1 inhibitor (50 µM Mdivi-1) and recombinant human HMGB1 (1 µg/mL) for 1 h prior to addition of OXP (10 µM) for 48 h. The caspase-3 activity was analyzed (n = 3). **p < 0.01. These data were obtained from three independent experiments, and the values represent the means ± S.D

Fig 5: Western blot analysis of protein expression of VCAM1, ICAM1, p65NF?B, SOD2, TXN1, HMGB1, and phospho-ERK1/2 after 4, 24, and 72 h exposure to different gas conditions. N, normoxia (21% O2); A, anoxia (0% O2); H, moderate hypoxia (10% O2); IH, intermittent hypoxia (0–21% O2). Images show representative blots, graphs depict densitometric quantifications from three samples.