Fig 2: Immunohistochemical staining of colonic mucosa with cyclooxygenase (COX) subtype specific antibodies in patients with colorectal neoplasia. a COX-2 immunohistochemical staining appears localized to the cytoplasm of absorptive cells (yellow arrows). No staining is seen in goblet cells (red arrows). A pericryptal stromal cell is marked (green arrow). b COX-1 immunohistochemical staining, here with antibody sc-1752, appears to stain morphological appearing tuft cells (red arrows) and in the lamina propria proposedly myofibroblasts (green arrow). c Fluorescence double labeling shows no co-localization between COX-1 (green) and specific markers for endocrine cells (red). Arrows point at COX-1 positive tuft cells. Subfigures 1, 2, 3, 5 and 7 have proportions of 100 µm per centimeter, subfigures 4 and 6 have proportions of 50 µm per centimeter. 1: Glucagon-like peptide-1, 2: Somatostatin, 3: Gastric Inhibitory Polypeptide, 4: Proprotein Convertase-1, 5: Proprotein Convertase-2, 6: Serotonin, 7: Chromogranin A

Fig 3: Microglial activation and PGD2 production observed in the hippocampus of WT and GPR120 KO mice. The protein level determined by western blot analysis (A). The level of Iba-1 protein in the hippocampus (B). The immunofluorescence of Iba-1 (C–E) and Iba-1 positive cell counts in the CA3 (D) and hippocampus (E). Scale bar = 50 µm. PGD2, PGE2, and PGF2a contents (F) in the hippocampus. PGD2 contents in each tissue (G). The level of COX-1 (H), COX-2 (I), L-PGDS (J), and H-PGDS (K) protein in the hippocampus. Data are means ± SEM, n = 5 per group. Statistical analysis was performed using unpaired Student's t test (*p < 0.05; **p < 0.01; ***p < 0.001 vs. WT)

Fig 4: High glucose causes a redox-sensitive induction of ECs senescence and promotes oxidative stress in ECs involving NADPH oxidase and cyclooxygenases, which is inhibited by empagliflozin. A, ECs are exposed to either normal or high glucose for 24 h in the presence or absence of empagliflozin (100 nmol/L), before dihydroethidium staining. Ethidium fluorescence is determined by confocal microscope. B, ECs are exposed either with N-acetyl cysteine (NAC, 1 mmol/L), VAS-2870 (VAS, a NADPH oxidase inhibitor, 5 µmol/L), or Indomethacin (INDO, a COX inhibitor, 30 µmol/L) for 30 min before the addition of high glucose for 96 h, and the subsequent determination of SA-ß-gal activity by flow cytometry. C, ECs are exposed to normal or a high glucose for 96 h in the presence or absence of empagliflozin (100 nmol/L) before Western blot analysis of the NADPH oxidase subunits p22phox, p47phox, COX-1 and COX-2. Results are shown as representative immunoblots (upper panels) and corresponding cumulative data (lower panels). Data are expressed as mean ± SEM of n = 4-5. *P < 0.05 vs. control normal glucose and # P < 0.05 vs. control high glucose

Fig 5: Immunoblot showing the intensity of apoptotic cascade components. (A) Western blotting analysis showing the intracellular expression of apoptotic cascade components in experimental groups of ARPE-19 cells. (1) ARPE-19 cells untreated, (2) ARPE-19 cells treated with 7.5 mM SI, (3) ARPE-19 cells simultaneously treated with 7.5 mM SI and 1µM of aB-wt, (4) ARPE-19 cells simultaneously treated with 7.5 mM SI and 1µM of aB?54–61. (B) Bar graphs of proteins of Bcl-2, EGR-1, and COX-1 normalized to cellular ß-actin protein levels. The asterisks (***) indicate a p-value < 0.005.