Fig 2: Analysis of the dynamics of HSP induction. Tissues were taken from 3- and 22-month-old mice 4 h after treatment with HSP990 (12 mg/kg) or vehicle. (A) qPCR analysis of the expression levels of Hspa1a/b, Dnajb1 and Hspb1 in brain regions and peripheral tissues. Values were calculated relative to vehicle-treated young mice. (B) Relative protein level of HSP70, HSP40 and HSP25 in the heart. Densitometric values were calculated relative to vehicle-treated young mice. (C) Representative western blots for HSP70, HSP40 and HSP25. Data are the mean ± SEM. 3 < n < 6/group. *P < 0.05; **P < 0.01; ***P < 0.001. Asterisk indicates the statistically significant difference in the level of induction. L.C., loading control; mth, months.

Fig 3: Comparison of HSP70 plasma levels (ng/ml) in SHAM and DJOS-operated groups subjected to the different dietary patterns pre- and post-surgery. Abbreviations: CD, control diet; HFS, high-fat, high-sugar diet. Vertical lines depict 95% confidence interval

Fig 4: DAMP release after (–)-Guaiol treatment in vitro. A The CRT level in three separate experiments detected by flow cytometry. B Representative image of CRT location and expression of three confocal images. Green, CRT; Red, β-catenin indicates the cell membrane; Blue, DAPI indicates the nucleus. C ATP content in the supernatant detected by an ATP assay kit. D Content of HMGB1 in the cell supernatant detected by enzyme-linked immunosorbent assay (ELISA). E Content of HSP70/90 in the cell supernatant detected by ELISAs. **p < 0.01, ***p < 0.001 vs. the control group

Fig 5: Effect of apoptosis inhibitor on (–)-Guaiol induced DAMP release. A Apoptosis detected by flow cytometry. B The CRT level detected by flow cytometry. C ATP content in the supernatant detected by an ATP assay kit. D Content of HMGB1 in the cell supernatant detected by enzyme-linked immunosorbent assays (ELISAs). E Content of HSP70/90 in the cell supernatant detected by ELISAs. **p < 0.01, ***p < 0.001 vs. the control group; #p < 0.01, ##p < 0.01, ###p < 0.001 vs. the (–)-Guaiol group