Fig 2: IHC for the protein expression in endometrial tissue in rats. (A, B) The expression of PTGFR: control (A), model (B). (C, D) The expression of TBXA2R: control (C), model (D). (E, F) The expression of COX-1: control (E), model (F). (G, H) The expression of COX-2: control (G), model (H). (I) % area for the immunohistochemical staining intensity in endometrial tissue in rats. Bars with different letters are statistically different (**P < 0.01). Values are means ± SD, n = 3 per group.

Fig 3: Over-expression of MEG8 attenuated hypoxia-induced VSMC proliferation, inflammation and migration.MEG8 overexpressing plasmids were transfected in A-10 cells or hVSMCs, which were then induced with hypoxia. (A) The MEG8 overexpression model was established, and the MEG8 level was detected by qRT-PCR. (B) Cell proliferation was verified by the CCK-8 assay. (C) Cell migration was determined by the Transwell assay. (D) Wound healing assay was employed to analyze cell migration. (E) The protein expression of MMP3, MMP9 and MMP13 in A-10 cells or hVSMCs was compared by Western blot. (F) The profiles of COX2, MIP-1ß, VCAM-1 and ICAM-1 in A-10 cells or hVSMCs were checked by Western blot. *p < 0.05, **p < 0.01, ***p < 0.001 (vs.NC group), #p < 0.05, ##p < 0.01, ###p < 0.001 (vs. hypoxia+NC group). All experiments were repeated three times, and each time was performed in triplicate.

Fig 4: Characterization of severe SAE and light SAE mice and dependence of sepsis-induced brain injury on gut microbiota. Survival rates of mice were observed until 7 days post-CLP. Mice that died within 48 hours of CLP were defined as severe SAE (SSAE) because of the high mortality. Mice that survived after 7 days were defined as light SAE (LSAE) because of the mortality rate tends to be flat. (A) Survival rate. (B) Neurobehavioral scores. (C) Representative H&E-stained sections of hippocampus of SSAE and LSAE mice. (D) Western blot analysis of iNOS, COX-2, and BDNF protein expressions in the hippocampus of SSAE and LSAE mice. GAPDH served as the loading control. Bar graphs depict the optical density of iNOS, COX-2, and BDNF expression. (E) Schematic illustration of FMT experiment: Mice were administered intragastric vancomycin (100 mg/kg), neomycin sulfate (200 mg/kg), metronidazole (200 mg/kg), and ampicillin (200 mg/kg) once-daily for 5 days to deplete the gut microbiota; subsequently, they received feces resuspended in PBS from SSAE and LSAE mice for 3 days. CLP surgery was performed, and mice were sacrificed 12 hours after CLP. (F) Representative H&E-stained sections of hippocampus of recipient mice. (G) Western blot analysis of iNOS, COX-2, and BDNF protein expressions in the hippocampus of recipient mice. GAPDH served as the loading control. Bar graphs depict the optical density of iNOS, COX-2, and BDNF expressions. Data presented as mean ± SD (n = 3–8 per group). Scale bars: 100µm and 20µm. *P < 0.05, **P < 0.01, ***P < 0.001 by Student’ t-test.

Fig 5: Increased mPGES1 protein abundance is dependent upon activation of NCC. Western blot (WB) analysis of PGE2 synthesis enzymes in tissue of kidney cortex. (A) mPGES1, total Cox2 (both bands), and Cox1 (B) quantification. Statistical significance evaluated by two-way ANOVA (see text), followed by Tukey’s post hoc tests (*p < 0.05).