Journal: Mediators of Inflammation

Article Title: Nobiletin Decreases Inflammatory Mediator Expression in Tumor Necrosis Factor-Stimulated Human Periodontal Ligament Cells

doi: 10.1155/2021/5535844

Figure Lengend Snippet: Effects of nobiletin on TNF-induced PTGS2 expression in HPDLCs. HPDLCs were stimulated by TNF (10 ng/ml) with or without nobiletin (12.5, 25, or 50 μ M) for 24 hours, and then PTGS2 expression was determined by Western blot analysis. Each photograph is representative of the results of 3 separate experiments. The quantification was performed using image analysis software. ∗ p < 0.05 compared to the TNF stimulation without nobiletin.

Article Snippet: Nobiletin (item number: 15421) and the antibody against PTGS2 (item number: 10112) were purchased from Cayman Chemical (Ann Arbor, MI, USA).

Techniques: Expressing, Western Blot, Software

Journal: International Journal of Molecular Sciences

Article Title: How Is Arachidonic Acid Metabolism in the Uterus Connected with the Immune Status of Red Deer Females ( Cervus elaphus L.) in Different Reproductive Stages?

doi: 10.3390/ijms24054771

Figure Lengend Snippet: mRNA expression of 5-LO ( A ), LTA4H ( B ), LTC4S ( C ), PTGS2 ( D ), PGES ( E ), PGFS ( F ), and PGIS ( G ) in uterine tissues (endometrium and myometrium) on 4th and 13th day of the estrous cycle, and in pregnancy and anestrus phase. Data were normalized against GAPDH for mRNA expression using GraphPad PRISM (Version 8.3.0). Each bar represents one experimental group with SEM. Statistical differences were analyzed withy two-way analysis (ANOVA) of variance followed by Tukey’s post hoc test. The lowest statistical significance was p < 0.05. Asterisks indicate statistical differences between endometrium and myometrium (* p < 0.05; ** p < 0.01; *** p < 0.001). Different letters indicate statistical differences ( p < 0.05) between the experimental reproductive stages throughout endometrium (a, b) and myometrium (A, B), respectively.

Article Snippet: Afterward, the proteins were transferred to 0.2 μm nitrocellulose membranes in transfer buffer by electroblotting for 1.5 h. The membranes were blocked in a 5% solution of skimmed milk with 1x TBS-T for 1.5 h at room temperature (RT), and then incubated overnight at 4 °C with specific primary antibodies for 5-LO (Cayman, 160402, 1:1000), LTA4H (Cayman, 160250, 1:1000), LTC4S (Biorbyt, orb580627, 1:1000), PTGS2 (Cayman, 160107, 1:1000), PGES (Cayman, 160140, 1:1000), PGFS (Cayman, 10007940, 1:1000), and PGIS (Cayman, 160640, 1:1000).

Techniques: Expressing

Journal: International Journal of Molecular Sciences

Article Title: How Is Arachidonic Acid Metabolism in the Uterus Connected with the Immune Status of Red Deer Females ( Cervus elaphus L.) in Different Reproductive Stages?

doi: 10.3390/ijms24054771

Figure Lengend Snippet: Protein expression of 5-LO ( A ), LTA4H ( B ), LTC4S ( C ), PTGS2 ( D ), PGES ( E ), PGFS ( F ), and PGIS ( G ) in uterine tissues (endometrium and myometrium) on 4th and 13th day of the estrous cycle, and in pregnancy and anestrus phase. Data were normalized against ACTB for protein expression using GraphPad PRISM (Version 8.3.0). Each bar represents one experimental group with SEM. Statistical differences were analyzed with two-way analysis (ANOVA) of variance followed by Tukey’s post hoc test. The lowest statistical significance was p < 0.05. Asterisks indicate statistical differences between endometrium and myometrium (* p < 0.1; *** p < 0.001). Different letters indicate statistical differences ( p < 0.05) between the experimental reproductive stages throughout endometrium (a, b) and myometrium (A, B), respectively.

Article Snippet: Afterward, the proteins were transferred to 0.2 μm nitrocellulose membranes in transfer buffer by electroblotting for 1.5 h. The membranes were blocked in a 5% solution of skimmed milk with 1x TBS-T for 1.5 h at room temperature (RT), and then incubated overnight at 4 °C with specific primary antibodies for 5-LO (Cayman, 160402, 1:1000), LTA4H (Cayman, 160250, 1:1000), LTC4S (Biorbyt, orb580627, 1:1000), PTGS2 (Cayman, 160107, 1:1000), PGES (Cayman, 160140, 1:1000), PGFS (Cayman, 10007940, 1:1000), and PGIS (Cayman, 160640, 1:1000).

Techniques: Expressing

Journal: International Journal of Molecular Sciences

Article Title: How Is Arachidonic Acid Metabolism in the Uterus Connected with the Immune Status of Red Deer Females ( Cervus elaphus L.) in Different Reproductive Stages?

doi: 10.3390/ijms24054771

Figure Lengend Snippet: List of monoclonal antibodies used for labeling lymphocytes for flow cytometry.

Article Snippet: Afterward, the proteins were transferred to 0.2 μm nitrocellulose membranes in transfer buffer by electroblotting for 1.5 h. The membranes were blocked in a 5% solution of skimmed milk with 1x TBS-T for 1.5 h at room temperature (RT), and then incubated overnight at 4 °C with specific primary antibodies for 5-LO (Cayman, 160402, 1:1000), LTA4H (Cayman, 160250, 1:1000), LTC4S (Biorbyt, orb580627, 1:1000), PTGS2 (Cayman, 160107, 1:1000), PGES (Cayman, 160140, 1:1000), PGFS (Cayman, 10007940, 1:1000), and PGIS (Cayman, 160640, 1:1000).

Techniques: Labeling, Cytometry, Sequencing, Amplification

Journal: International Journal of Molecular Sciences

Article Title: How Is Arachidonic Acid Metabolism in the Uterus Connected with the Immune Status of Red Deer Females ( Cervus elaphus L.) in Different Reproductive Stages?

doi: 10.3390/ijms24054771

Figure Lengend Snippet: Oligonucleotide sequences used for real-time PCR. GAPDH—Glyceraldehyde 3-phosphate dehydrogenase, RN18S1—18S ribosomal RNA, ACTB—β-actin, 5-LO—5-Lipoxygenase, LTA4H—Leukotriene A4 hydrolase, LTC4S—Leukotriene C4 Synthase, PTGS2—prostaglandin endoperoxide synthase 2, PGES—Prostaglandin E2 synthase, PGFS—Prostaglandin F2alpha synthase, PGIS—Prostaglandin I2 synthase.

Article Snippet: Afterward, the proteins were transferred to 0.2 μm nitrocellulose membranes in transfer buffer by electroblotting for 1.5 h. The membranes were blocked in a 5% solution of skimmed milk with 1x TBS-T for 1.5 h at room temperature (RT), and then incubated overnight at 4 °C with specific primary antibodies for 5-LO (Cayman, 160402, 1:1000), LTA4H (Cayman, 160250, 1:1000), LTC4S (Biorbyt, orb580627, 1:1000), PTGS2 (Cayman, 160107, 1:1000), PGES (Cayman, 160140, 1:1000), PGFS (Cayman, 10007940, 1:1000), and PGIS (Cayman, 160640, 1:1000).

Techniques: Real-time Polymerase Chain Reaction

Journal: American journal of therapeutics

Article Title: Pilot Study of Tart Cherry Juice for the Treatment of Insomnia and Investigation of Mechanisms

doi: 10.1097/MJT.0000000000000584

Figure Lengend Snippet: A. Relative induction of indoleamine 2,3-dioxygenase (IDO) and COX-2 in Caco-2 cells stimulated with IFN-γ and decrease of IDO and COX-2 in cells treated with procyanidin B-2. B. Relative induction of NF-KB in Caco-2 cells stimulated with IFN-γ. 1= control, 2= IFN-γ, 3= 12.5 μM procyanidin B-2, 4= 25 μM procyanidin B-2, and 5= 50 μM procyanidin B-2

Article Snippet: Primary antibody against cyclooxygenase-2 (COX-2) was purchased from Cayman Chemical (Ann Arbor, MI).

Techniques:

Journal: Nature Communications

Article Title: Sialic acid exacerbates polycystic ovary syndrome in mice by modulating gut microbiota-mediated bile acid metabolism and FXR activation

doi: 10.1038/s41467-026-71365-4

Figure Lengend Snippet: A , B Representative images of TUNEL-stained ovarian sections (scale bar: 50 μm). C , D Representative Western blot images of PTGS2 and GPX4 in ovarian tissues. Band quantification is normalized to the first lane. E KGN cells were pretreated with 1 μM ferrostatin-1 (Fer-1) with or without 0.1 μg/mL rMuIL-22 for 2 h, followed by DHEA (20 μM) treatment for 24 h. Cell viability was assessed using the CCK-8 assay ( n = 5 biological replicates per group). F Representative H&E-stained ovarian tissue sections (scale bar: 200 μm). G Quantification of cystic follicles and corpora lutea ( n = 5 mice per group). H Representative Western blot images of p-STAT3 and total STAT3 in ovarian tissues of rMuIL-22-treated mice. Band quantification is normalized to the first lane (p-STAT3/STAT3). I , J KGN cells were treated with 20 μM Stattic for 1 h, followed by rMuIL-22 (0.1 μg/mL) for 2 h, and then DHEA (20 μM) for 24 h. I Cell viability ( n = 6 biological replicates per group). J Representative images of PTGS2 and GPX4. Band quantification is normalized to the first lane. K GTT and ITT assays ( n = 5 mice per group). GTT: ## p = 0.0097 (15 min), ### p = 0.0006 (30 min), ## p = 0.0019 (60 min) and ## p = 0.0046 (90 min) for rMuIL-22 + Stattic + Neu5Ac + DHEA vs rMuIL-22 + Neu5Ac + DHEA. ITT: # p = 0.0412 (60 min) for rMuIL-22 + Stattic + Neu5Ac + DHEA vs rMuIL-22 + Neu5Ac + DHEA. L Representative H&E-stained images (scale bar: 200 μm). M Quantification of cystic follicles and corpora lutea ( n = 5 mice per group). Data are presented as mean ± SD. One-way ANOVA followed by Tukey’s post hoc test ( E , G , I , and M ), and two-way repeated-measures ANOVA with Sidak’s multiple-comparisons correction ( K ) were performed. TUNEL-stained and Western blot images are representative of at least three biologically independent experiments with consistent results ( A – D , H , and J ).

Article Snippet: Subsequently, the following primary antibodies were applied for overnight incubation at 4 °C: PTGS2 (1:1000, AF7003, Affinity Biosciences, USA), GPX4 (1:1000, AF7003, Affinity Biosciences, USA), STAT3 (1:1000, AF6294, Affinity Biosciences, USA), p-STAT3 (1:1000, AF3293, Affinity Biosciences, USA), and β-actin (1:1000, AF7018, Affinity Biosciences, USA).

Techniques: TUNEL Assay, Staining, Western Blot, CCK-8 Assay