Fig 2: (A) is the representative immunofluorescence image of paraventricular nucleus (PVN) in a rat, marking the location of PVN on both sides of the third ventricle (400 HPF), bar = 30 µm. The blue dots represented the nucleus. (B) is representative image of multiplex immunofluorescence staining from the sham + shCtrl, sham + shTLR4, MI + shCtrl and MI + shTLR4 groups of TLR4, MyD88 and NF‐kB (400 HPF) in the PVN, bar = 30 µm. (C) is representative image of multiplex immunofluorescence staining from the sham and MI groups of TLR4, MyD88 and NF‐kB (400 HPF) in the PVN, bar = 15 µm. The number of TLR4‐, MyD88‐, NF‐kB‐ and nuclei‐positive cells in each group was counted and is presented as the mean ± SD (D‐G). n = 5 in each group. *p < 0.05 compared with sham group. # p < 0.05 compared with MI + shCtrl. Abbreviations: MI, myocardial infarction; PVN, paraventricular nucleus; shCtrl, control virus; shTLR4, TLR4‐inhibiting virus; 3V, the third ventricle

Fig 3: Amniotic epithelial cells expressed a functional RAGE axis. (A) RNA expressions of RAGE, TIRAP, Myd88, and Diaphanous-1 were detected by RT-PCR in pAECS. Negative controls were performed in the absence of a cDNA template. (B) The RAGE receptor and its adaptors, Diaphanous-1, MyD88, and TIRAP (green staining, Alexa488), were detected by immunocytochemistry on primary amniocytes (pAECs). Nuclei were counterstained with Hoechst (blue). Scales bars: 50 µM (magnification ×200). Negative control was realized without primary antibody hybridization.

Fig 4: Effect of ATP2B1-AS1 and miR-330-5p on the cell viability, apoptosis, cytokines, and TLR4-MyD88-NF-κB signaling pathway in OGD/R PC12 cells. (A–D) The effect of pcDNA3.1-ATP2B1-AS1 and miR-330-5p mimics on the cell viability (A), LDH level (B), Caspase 3 activity (C), and apoptosis rate (D) of OGD/R PC12 cells. PcDNA3.1-ATP2B1-AS1 reduced the cell viability, increased the LDH level, Caspase 3 activity, and apoptosis rate of OGD/R PC12 cells. These effects of ATP2B1-AS1 on OGD/R PC12 cells were abrogated by miR-330-5p mimics. *P < 0.05, **P < 0.01, ***P < 0.001.

Fig 5: Altered PBMC Protein Expression with acute high‐fat feeding. Normalized and fold change results show PBMC RAGE (A), ADAM10 (B), TLR4 (C), and MyD88 (D) with acute high‐fat feeding with and without prior aerobic exercise. Western blot bands follow the same order as the figure bars. Data were analyzed via two‐way ANOVA and data are presented as mean ± SEM. n = 12 for all proteins represented. ***Main effect of HFM P = 0.001.