Cox1 Antibody - #AF7002

FIGURE 2 Effect of Urolithin A on mitochondria‐related genes in SACI mice. (A) Venn diagram of gene expression in sham group vs. SACI group and SACI group vs. UA group. (B) Heatmap analysis of 90 overlapping gene expressions in three groups (n = 4). (C) GO term analysis of 90 overlapping gene expressions in the SACI group vs. UA group. (D) Volcano plots of differential genes in the sham group vs. SACI group and UA group vs. SACI group. (E) Western blotting and quantitative analysis of COX I, COX IV, TOMM7, and NDUFC1 in cardiac tissues of the sham, SACI, and UA groups (n = 3). (F) Representative transmission electron microscopy (TEM) images of cardiac tissues in the sham, SACI, and UA groups (scale bar = 2 μm; 500 nm). (G) Mitochondrial membrane potential changes in isolated purified heart mitochondria in the sham, SACI, and UA groups. (JC‐1 fluorescence: red/green ratio; n = 4). (H) Changes in total ATP concentration in heart tissues in the sham, SACI, and UA groups (n = 6). The results are presented as the mean ± SD. **p < 0.01, ***p < 0.005, ****p < 0.0001 vs. sham group; ##p < 0.01, ###p < 0.005, ####p < 0.0001 vs. SACI group, by one‐way ANOVA tests followed by Tukey tests.

Figure 2 Comparison of COX-1 levels: Microscopic photography (a) control group (Con-Run, 3); (b) bee-pollen-supplemented group (BP-Run; 4). In both pictures, cytoplasmatic reaction seems to be comparable, although in the control group, there seem to be more deposits of antibodies linked to COX-1 around the nuclei of gastric cells, resulting in their darker tone and classification as cells with increased COX-1 levels.

FIGURE 1 Aspirin inhibits myocardial inflammation and alleviates cardiac dysfunction in response to I/R injury. (A) Representative electrocardiogram shows the successful establishment of myocardial I/R injury model (n = 5 per group). The red arrows represent changes in the ST segment. (B) Representative images show TTC staining of the myocardium from rats (n = 5 per group) belonging to the sham, I/R, and I/R + 50 mg/kg aspirin groups. The infarct size quantification of the 3 groups of rats is also shown. (C) ELISA assay results show the serum levels of creatine kinase-MB (CK-MB) and cardiac troponin T (cTnT) in the sham, I/R, and I/R + aspirin group rats (n = 5 per group). (D) Laggendorff in vitro cardiac perfusion results show the left ventricular developed pressure (LVDP), maximum and minimum rates of pressure development (max dP/dt and min dP/dt), and heart rate (HR) in the sham, I/R, and I/R + aspirin group rats (n = 5 per group). (E) Representative H&E staining images (×200 and ×400) show the histological details of the myocardium from the sham, I/R, and I/R + aspirin group rats (n = 5 per group). (F,G) Representative immunoblots and quantification of the expression levels of COX-1 and COX-2 in the sham, I/R, and I/R + aspirin group rats (n = 5 per group). (H) ELISA results show the tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) level in the myocardial tissues of the sham, I/R, and I/R + aspirin group rats (n = 5 per group). The tissues were harvested 2 h post-reperfusion. The data are represented as means ± SD; *p < 0.05 vs. the sham group; # p < 0.05 vs. the I/R group.

FIGURE 3 Gasrtodin alleviates dual gastric mucosal injury by suppressing inflammation. (A) Representative images of the harvest stomach specimens from the sham, I/R, I/R + aspirin, I/R + 50 mg/kg aspirin + 200 mg/kg gastrodin group, and I/R + 200 mg/kg gastrodin group rats (n = 5 per group). (B) Byron gastric mucosa injury scores of the sham, I/R, I/R + aspirin, I/R + aspirin + gastrodin group, and I/R + gastrodin group rats (n = 5 per group). The scoring criteria are shown in Table 1. (C) Representative images (×100 and ×200) of the H&E-staining gastric tissue sections from the sham, I/R, I/R + aspirin, I/R + aspirin + gastrodin group, and I/R + gastrodin group rats (n = 5 per group). (D) Representative immunofluorescence staining images (×200) show the COX-1 and COX-2 expression levels in the gastric tissues of the sham, I/R, I/R + aspirin, I/R + aspirin + gastrodin group, and I/R + gastrodin group rats (n = 5 per group). The green denotes COX-1, red denotes COX-2, and blue denotes the nucleus. (E–G) ELISA assay results show the levels of cyclooxygenase-1 (COX-1), cyclooxygenase-2 (COX-2), tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), prostaglandin E2 (PGE2) and endothelin (ET) in the gastric tissues of the sham, I/R, I/R + aspirin, I/R + aspirin + gastrodin group,and I/R + gastrodin group rats (n = 5 per group). The data are represented as means ± SD; * p < 0.05 vs. the sham group; # p < 0.05 vs. the I/R group; ^ p < 0.05 vs. the I/R + aspirin group; & p < 0.05 vs. the I/R + aspirin + gastrodin group.

FIG 6 FbaA initiates autophagy upon Fn-mediated binding to integrin 51. (A and B) Western blot analysis of TLR2 (A) and TLR4 (B) expression in Hep2 cells stimulated with FbaA for the indicated times. (C and D) Expression of LC3II in Hep2 cells with and without TLR2- or TLR4-specific siRNA following FbaA stimulation. (E) Schematic representation of the Fn subunit. Fn contains domain types I, II, and III. FnI is the region critical for binding to FnBP; FnIII9 and FnIII10 contain motifs that interact with integrins. (F) Hep2 cells were transfected with GST-FbaA, GST-FbaAFn, or GST (as a control), and the integrin 5 or 1 chain was measured after being pulled down with anti-GST Ab. (G) Expression of Fn, integrin 5, integrin 1, and LC3II in Hep2 cells stimulated with FbaA at the indicated concentrations, as analyzed by Western blotting. (H) Western blot analysis of LC3II expression in Hep2 cells stimulated with different concentrations of Fn and mock-FbaA or FbaA. (I to K) Expression of LC3II in Hep2 cells with or without specific siRNA against Fn, integrin 5, or integrin 1 following FbaA stimulation, as analyzed by Western blotting. (L and M) Fn, integrin 5, and integrin 1 knockdown cells were pretreated without (L) or with (M) rapamycin for 0.5 h and then infected with M1 GAS strain SF370 for 6 h. Numbers of CFU of M1 GAS strain SF370 were counted in the cells. (N) M1 GAS strain SF370-infected sh-Atg5-Hep2 cells with specific siRNA against Fn, integrin 5, or integrin 1. Numbers of CFU of M1 GAS strain SF370 were counted in the cells. *, P  0.05.

FIG 6 |FbaA initiates autophagy upon Fn-mediated binding to integrin 51. (A and B) Western blot analysis of TLR2 (A) and TLR4 (B) expression in Hep2 cells stimulated with FbaA for the indicated times.