Fig 2: Effects of LPS on heart tissue structure and inflammation response marker levels. (A) Western blot analysis was performed to evaluate the levels of PAI-1 and TNF-a in the heart tissues after 1, 2, 3 and 4 days of LPS injection. GAPDH was used as a loading control. (B) Histopathology of heart tissues with or without LPS injection for 4 days was assessed by staining tissues with hematoxylin and eosin under a light microscope. scale bar, 100 µm. (C) Western blot analysis was performed to evaluate the levels of IL-6, I?Ba and p-I?Ba in the heart tissues with or without LPS injection for 4 days. GAPDH was used as a loading control. *P<0.05, **P<0.01, ***P<0.001 vs. the control group. LPS, lipopolysaccharide; PAI-1, plasminogen activator inhibitor 1; TNF-a, tumor necrosis factor-a; HE, Hematoxylin eosin; IL-6, interleukin 6; I?Ba, inhibitor of nuclear factor ?B kinase a; p, phosphorylated.

Fig 3: LPS-regulated transcriptome profile in heart. (A) Heat map representing the genes with expression levels that were altered after 4 days of LPS treatment in the mouse heart. Gene expression is shown with a pseudocolor scale, where red denotes higher gene expression levels and green denotes lower gene expression levels (P<0.05). (B) RT-qPCR was performed to verify the expression levels of FZD8, IL-8, PAI-1 and FGF-21, and the data were compared with the RNA-sequencing data. GAPDH was used as the internal control. *P<0.05 vs. the control group. LPS, lipopolysaccharide; RT-qPCR, reverse transcription-quantitative PCR; FZD8, frizzled class receptor 8; IL-8, interleukin 8; PAI-1, plasminogen activator inhibitor 1; FGF21, fibroblast growth factor 21.

Fig 4: Rb1 reduced cellular adhesion molecule expression in aged mouse thoracic aortas. Western blot analysis of ICAM-1, VCAM-1 and PAI-1 expression in thoracic aorta tissues. The data are expressed as the mean ± SD. **p < 0.01 vs. the Young group; #p < 0.05, ##p < 0.01 vs. the Old + Vehicle group.

Fig 5: Blocking TIMP1, PAI1 and IGFBP1 affects IL12a-mediated tube formation. HUVECs suspended in CM were seeded into Matrigel-coated 96-well plates. Sum of length of the trees composed from segments and branches in the analyzed area (Tot. branching length) were analyzed by usingimage jsoftware. The unit of length is pixel. (A) Supernatants from pc-IL12A HGC27 cells pretreated with PAI1, IGFBP1 or control neutralizing antibodies (Ctrl) for 6 h were used as the CM. (B) Supernatants from sh-IL12A AGS cells pretreated with TIMP1 or control neutralizing antibodies (Ctrl) for 6 h were used as the CM. Data are shown as the mean ± SD; statistical test: Student’st-test and one-way ANOVA with the Dunnettpost hoctest;n = 3; *P < 0.05, **P < 0.01, ***P < 0.001. Scale bars, 300 µm.