Journal: The Journal of clinical endocrinology and metabolism

Article Title: Prostaglandin E2 as a regulator of germ cells during ovarian development.

doi: 10.1210/jc.2009-0755

Figure Lengend Snippet: FIG. 1. A, Representative nonquantitative RT-PCR analysis of COX1, COX2, PTGES, EP2, and EP4 mRNA from 64-d (1T), 14-wk (14), and 18-wk (18) human fetal ovary or endometrium (E) cDNA as indicated. Plus and minus signs indicate the presence or absence of reverse transcriptase during cDNA synthesis. M, 100bp ladder marker; bl, water blank. GAPDH was analyzed as a positive control. B, Quantitative RT-PCR analysis of mRNA expression of COX1, PTGES, EP2, and EP4 in human ovary at different gestations. *, P 0.05; **, P 0.01. Mean SEM of n 4–10 per group. Quantities are expressed as percentage relative to GAPDH.

Article Snippet: Secondary antibodies were swine antirabbit biotinylated (E0353), diluted 1:500 (EP2 and EP4); goat antirabbit biotinylated (E0437), diluted 1:500 (PTGES) (both from Dako, Cambridge, UK); or chicken antigoat biotinylated (sc-2984) (Santa Cruz Biotechnology) diluted 1:200 (COX1 and COX2).

Techniques: Reverse Transcription Polymerase Chain Reaction, Reverse Transcription, cDNA Synthesis, Marker, Positive Control, Quantitative RT-PCR, Expressing

Journal: The Journal of clinical endocrinology and metabolism

Article Title: Prostaglandin E2 as a regulator of germ cells during ovarian development.

doi: 10.1210/jc.2009-0755

Figure Lengend Snippet: FIG. 3. Immunohistochemical localization of EP2 (A–C, H, and I) and EP4 (D–F, G, and I) in second-trimester human fetal ovaries. Gestations: A, D, and E, 14 wk; G–I, 17 wk; B and F, 19 wk; and C, 20 wk. Insets show peptide-blocked negative controls. A–F, Positive 3,3-diaminobenzidine tetrahydrochloride staining is brown, and sections are counterstained with hematoxylin. S1, Somatic cell within “cord”; S2, somatic cell among germ cells; gc, germ cell; PF, primordial follicle. Scale bars are 50 m. G–I, Dual immunofluorescence for EP2 (red, H and I) and EP4 (green, G and I). Counterstain is DAPI (blue, I). Separate channel (G and H) and merged (I) images are shown. Yellow in I indicates colocalization, although the intensity of the red staining tends to swamp out the green staining (compare I and G). Scale bars are 20 m.

Article Snippet: Secondary antibodies were swine antirabbit biotinylated (E0353), diluted 1:500 (EP2 and EP4); goat antirabbit biotinylated (E0437), diluted 1:500 (PTGES) (both from Dako, Cambridge, UK); or chicken antigoat biotinylated (sc-2984) (Santa Cruz Biotechnology) diluted 1:200 (COX1 and COX2).

Techniques: Immunohistochemical staining, Staining, Immunofluorescence

Journal:

Article Title: Differential Regulation of The Aggressive Phenotype of Inflammatory Breast Cancer Cells By Prostanoid Receptors EP3 and EP4

doi: 10.1002/cncr.25167

Figure Lengend Snippet: Figure 2A. Effect of PGE2 and EP4 Agonist PGE1-Alcohol on Invasion. PGE2 and the EP4 agonist PGE1-alcohol induced significantly increased invasion of SUM149 cells but had no effect on invasion of MDA-MB-231 cells.

Article Snippet: The shRNA constructs against PTGER4 (EP4) were obtained from OriGene (Rockville, MD).

Techniques:

Journal: Arthritis research & therapy

Article Title: Pharmacological characterisation of CR6086, a potent prostaglandin E 2 receptor 4 antagonist, as a new potential disease-modifying anti-rheumatic drug.

doi: 10.1186/s13075-018-1537-8

Figure Lengend Snippet: Fig. 1 Biochemical characterisation of CR6086 activity on prostaglandin E2 receptor 4 (EP4) receptors. a Effect of CR6086 on prostaglandin E2 (PGE2) binding to EP4 receptor. Membranes from human embryonic kidney cells 293 (HEK293) cells transfected with human EP4 receptor were incubated with [3H]PGE2 and CR6086 (range 1–3000 nM). The results are expressed as percentage inhibition of specific binding. Data are representative of five independent experiments. b Effect of CR6086 on cyclic adenosine monophosphate (cAMP) release from membranes of HEK293 cells transfected with human EP4 receptor and stimulated with 3 nM PGE2 in the presence of CR6086 (range 3–300 nM). Data are representative of three independent experiments. c CR6086 inhibition affinity constant [Ki], and [3H]PGE2 dissociation constant [Kd] towards human and rodent EP4 receptors

Article Snippet: For rodent EP4 receptor cloning, the cDNA encoding the full-length rat (NM_032076) or mouse (NM_001136079) EP4 receptors cloned in pCMV6 entry was provided by OriGene Inc. (TrueORF Gold, catalogue numbers RN206839 and MC216813; OriGene, Rockville, MD, USA).

Techniques: Activity Assay, Binding Assay, Transfection, Incubation, Inhibition

Journal: Arthritis research & therapy

Article Title: Pharmacological characterisation of CR6086, a potent prostaglandin E 2 receptor 4 antagonist, as a new potential disease-modifying anti-rheumatic drug.

doi: 10.1186/s13075-018-1537-8

Figure Lengend Snippet: Fig. 5 Pharmacokinetics of CR6086 in rats. a Unbound concentrations of CR6086 administered orally in the dose range of 1–30 mg/kg. The dashed line represents the inhibition constant (36.2 ng/ml) value for the rat prostaglandin E2 receptor 4. b Pharmacokinetic parameters

Article Snippet: For rodent EP4 receptor cloning, the cDNA encoding the full-length rat (NM_032076) or mouse (NM_001136079) EP4 receptors cloned in pCMV6 entry was provided by OriGene Inc. (TrueORF Gold, catalogue numbers RN206839 and MC216813; OriGene, Rockville, MD, USA).

Techniques: Drug discovery, Inhibition

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: EP4 receptor stimulation in combination with core decompression therapy enhanced bone regeneration in a canine model of osteonecrosis of femoral head

doi: 10.3389/fbioe.2025.1622918

Figure Lengend Snippet: HE staining and immunohistochemistry of human femoral heads from patients with hip fracture (upper panels, 66 years old, female) and ONFH (lower panels, 73 years old, female). The black boxes are shown at higher magnification. Arrowheads indicate EP4-positive lining cells. HE, hematoxylin–eosin; ONFH, osteonecrosis of the femoral head.

Article Snippet: Immunohistochemical staining for EP4 was performed using a rabbit anti-EP4 antibody (1.0 mg/mL, NLS3890; Novus Biologicals, Centennial, CO, United States) or rabbit IgG (I-1000; Vector Laboratories, Newark, CA, United States) as a negative control.

Techniques: Staining, Immunohistochemistry

Journal: Hypertension research : official journal of the Japanese Society of Hypertension

Article Title: Role of angiotensin-converting enzyme 2/angiotensin-(1-7)/Mas axis in the hypotensive effect of azilsartan.

doi: 10.1038/hr.2014.49

Figure Lengend Snippet: Figure 1 Effect of azilsartan or olmesartan on systolic (a) and diastolic (b) blood pressure in hRN/hANG-Tg mice. n ¼ 5 for each group. *Po0.01 vs untreated hRN/hANG-Tg. **Po0.01 vs the same dose in olmesartan- treated mice

Article Snippet: Then, hRN/hANG-Tg mice were separated into two groups receiving either 1 mg kg 1 per day of azilsartan (provided by Takeda, Tokyo, Japan) or olmesartan (Olmesartan Acid, Toronto Research Chemicals, Toronto, Ontario, Canada) in chow for 2 weeks.

Techniques: