Journal: BMC Cancer

Article Title: Possible predisposition for colorectal carcinogenesis due to altered gene expressions in normal appearing mucosa from patients with colorectal neoplasia

doi: 10.1186/s12885-019-5833-8

Figure Lengend Snippet: Simplified schematic illustration of pathways for Wnt/β-catenin, ERK/MAPK and PI3K/Akt and PGE 2 -metabolism. A) Canonical Wnt/β-catenin signaling. The engagement of the Wnt receptor, Frizzled, leads to the inhibition of the β-catenin destruction complex, composed of APC, axin and GSK3β. β-catenin thereby avoids ubiquitination and subsequent degradation, thus allowing it to translocate to the nucleus to activate an array of regulatory genes. B) The RAS/RAF/MEK/ERK MAPK pathway. Stimulation of the receptor tyrosine kinase (RTK) or G-protein coupled receptors (GPCRs) leads to sequential activation of RAS, RAF, MEK, and ERK causing modification of substrates promoting cell survival and proliferation. C) In the PI3K/Akt pathway, activation of the RTK or GPCRs leads to sequential modification of phosphatidyl inositol residues of the phospholipid bilayer. In this process, PI3K generates PIP3. PIP3 in association with PDK1 activates Akt. Akt then modulates the activity of downstream substrates including mTOR, thus promoting proliferation and cell survival. D) PGE 2 -metabolism. PGE 2 -synthesis begins with catalytic hydrolysis of membrane phospholipids by cytoplasmic phospholipase A2 (cPLA 2 ), thus releasing arachidonic acid (AA). By the action of the COX-1 and COX-2, AA is converted to prostaglandin H 2 (PGH 2 ). PGH 2 is then converted to PGE 2 by prostaglandin E synthase (PTGES). The main exporter of PGE 2 is thought to be multi-drug resistance related polypeptide 4 (MRP4). Removal of PGE 2 from the extracellular compartment around target cells occurs by diffusion to the blood stream and subsequent uptake and degradation in lung, liver or kidney endothelial cells or by import to colonic epithelial cells through the prostaglandin transporter (PGT) and subsequent degradation by 15-prostaglandin dehydrogenase (15-PGDH). Through autocrine and paracrine signaling, extracellular PGE 2 stimulates the prostaglandin receptors EP1–4. The EPs are GPCRs with EP1 being Gαq-coupled while EP2 and EP4 are Gαs-coupled. EP3 is capable of coupling with different G-proteins including Gαi, Gαs and Gαq

Article Snippet: The following antibodies were tested: Mouse anti-EP3 antibody (5F5), sc-57,105 (Santa Cruz Biotechnology), Rabbit anti-EP3 antibody, 101,760 (Cayman Chemicals, Ann Arbor, MI, USA), Rabbit anti-EP3 antibody, APR-065 (Alomone Labs, Jerusalem, Israel), Rabbit anti-EP3 antibody (PTGER3), NBP1–84835 (Bio-techne, Abingdon, United Kingdom).

Techniques: Inhibition, Activation Assay, Modification, Activity Assay, Diffusion-based Assay

Journal: BMC Cancer

Article Title: Possible predisposition for colorectal carcinogenesis due to altered gene expressions in normal appearing mucosa from patients with colorectal neoplasia

doi: 10.1186/s12885-019-5833-8

Figure Lengend Snippet: Quantitative real-time PCR primer sequences

Article Snippet: The following antibodies were tested: Mouse anti-EP3 antibody (5F5), sc-57,105 (Santa Cruz Biotechnology), Rabbit anti-EP3 antibody, 101,760 (Cayman Chemicals, Ann Arbor, MI, USA), Rabbit anti-EP3 antibody, APR-065 (Alomone Labs, Jerusalem, Israel), Rabbit anti-EP3 antibody (PTGER3), NBP1–84835 (Bio-techne, Abingdon, United Kingdom).

Techniques: Real-time Polymerase Chain Reaction

Journal: BMC Cancer

Article Title: Possible predisposition for colorectal carcinogenesis due to altered gene expressions in normal appearing mucosa from patients with colorectal neoplasia

doi: 10.1186/s12885-019-5833-8

Figure Lengend Snippet: Difference in mRNA expression in CRN versus controls

Article Snippet: The following antibodies were tested: Mouse anti-EP3 antibody (5F5), sc-57,105 (Santa Cruz Biotechnology), Rabbit anti-EP3 antibody, 101,760 (Cayman Chemicals, Ann Arbor, MI, USA), Rabbit anti-EP3 antibody, APR-065 (Alomone Labs, Jerusalem, Israel), Rabbit anti-EP3 antibody (PTGER3), NBP1–84835 (Bio-techne, Abingdon, United Kingdom).

Techniques: Expressing

Journal: Biology of reproduction

Article Title: Immunolocalization of adipocytes and prostaglandin E2 and its four receptor proteins EP1, EP2, EP3, and EP4 in the caprine cervix during spontaneous term labor.

doi: 10.1095/biolreprod.111.096040

Figure Lengend Snippet: FIG. 1. Specificity of rabbit polyclonal anti-human EP receptor antibodies and perilipin A antibody in intrapartum caprine cervix. EP receptor and perilipin A expression in caprine cervix was assayed by Western blot analysis as described in Materials and Methods. Western blots demonstrated only a single apparent band with antibodies against EP1, EP2, EP4, and perilipin A, and two bands with the anti-EP3 antibody. Preabsorption of the primary antibodies with the related blocking peptide (BP) (þ signifies BP added) abolished the specific signals.

Article Snippet: Membrane proteins were blocked with Blotto A before Western blotting with primary rabbit polyclonal antibodies against human EP1, EP2, EP3, and EP4 as well as perilipin A and a compatible secondary HRP-linked goat anti-rabbit antibody (sc-2004; Santa Cruz Biotechnology, Inc.).

Techniques: Expressing, Western Blot, Blocking Assay

Journal: Biology of reproduction

Article Title: Immunolocalization of adipocytes and prostaglandin E2 and its four receptor proteins EP1, EP2, EP3, and EP4 in the caprine cervix during spontaneous term labor.

doi: 10.1095/biolreprod.111.096040

Figure Lengend Snippet: FIG. 7. Immunolocalization of the contractile EP3 receptor subtype in caprine uterine cervical tissues during parturition, nonpregnancy, and pregnancy. Labeling of EP3 receptor protein at parturition is shown in the cervical stroma (a), circular muscle layer (b), and canal epithelium (c), and during pregnancy and nonpregnancy in the glandular (e, h) and canal (d, g) epithelia, blood vessels (f), and stroma (e). No immunostaining is observed in the controls (i). Original magnification 3400; bar ¼ 20 lm.

Article Snippet: Membrane proteins were blocked with Blotto A before Western blotting with primary rabbit polyclonal antibodies against human EP1, EP2, EP3, and EP4 as well as perilipin A and a compatible secondary HRP-linked goat anti-rabbit antibody (sc-2004; Santa Cruz Biotechnology, Inc.).

Techniques: Labeling, Immunostaining

Journal: BMC Cancer

Article Title: Possible predisposition for colorectal carcinogenesis due to altered gene expressions in normal appearing mucosa from patients with colorectal neoplasia

doi: 10.1186/s12885-019-5833-8

Figure Lengend Snippet: Simplified schematic illustration of pathways for Wnt/β-catenin, ERK/MAPK and PI3K/Akt and PGE 2 -metabolism. A) Canonical Wnt/β-catenin signaling. The engagement of the Wnt receptor, Frizzled, leads to the inhibition of the β-catenin destruction complex, composed of APC, axin and GSK3β. β-catenin thereby avoids ubiquitination and subsequent degradation, thus allowing it to translocate to the nucleus to activate an array of regulatory genes. B) The RAS/RAF/MEK/ERK MAPK pathway. Stimulation of the receptor tyrosine kinase (RTK) or G-protein coupled receptors (GPCRs) leads to sequential activation of RAS, RAF, MEK, and ERK causing modification of substrates promoting cell survival and proliferation. C) In the PI3K/Akt pathway, activation of the RTK or GPCRs leads to sequential modification of phosphatidyl inositol residues of the phospholipid bilayer. In this process, PI3K generates PIP3. PIP3 in association with PDK1 activates Akt. Akt then modulates the activity of downstream substrates including mTOR, thus promoting proliferation and cell survival. D) PGE 2 -metabolism. PGE 2 -synthesis begins with catalytic hydrolysis of membrane phospholipids by cytoplasmic phospholipase A2 (cPLA 2 ), thus releasing arachidonic acid (AA). By the action of the COX-1 and COX-2, AA is converted to prostaglandin H 2 (PGH 2 ). PGH 2 is then converted to PGE 2 by prostaglandin E synthase (PTGES). The main exporter of PGE 2 is thought to be multi-drug resistance related polypeptide 4 (MRP4). Removal of PGE 2 from the extracellular compartment around target cells occurs by diffusion to the blood stream and subsequent uptake and degradation in lung, liver or kidney endothelial cells or by import to colonic epithelial cells through the prostaglandin transporter (PGT) and subsequent degradation by 15-prostaglandin dehydrogenase (15-PGDH). Through autocrine and paracrine signaling, extracellular PGE 2 stimulates the prostaglandin receptors EP1–4. The EPs are GPCRs with EP1 being Gαq-coupled while EP2 and EP4 are Gαs-coupled. EP3 is capable of coupling with different G-proteins including Gαi, Gαs and Gαq

Article Snippet: The following antibodies were tested: Mouse anti-EP3 antibody (5F5), sc-57,105 (Santa Cruz Biotechnology), Rabbit anti-EP3 antibody, 101,760 (Cayman Chemicals, Ann Arbor, MI, USA), Rabbit anti-EP3 antibody, APR-065 (Alomone Labs, Jerusalem, Israel), Rabbit anti-EP3 antibody (PTGER3), NBP1–84835 (Bio-techne, Abingdon, United Kingdom).

Techniques: Inhibition, Activation Assay, Modification, Activity Assay, Diffusion-based Assay

Journal: BMC Cancer

Article Title: Possible predisposition for colorectal carcinogenesis due to altered gene expressions in normal appearing mucosa from patients with colorectal neoplasia

doi: 10.1186/s12885-019-5833-8

Figure Lengend Snippet: Quantitative real-time PCR primer sequences

Article Snippet: The following antibodies were tested: Mouse anti-EP3 antibody (5F5), sc-57,105 (Santa Cruz Biotechnology), Rabbit anti-EP3 antibody, 101,760 (Cayman Chemicals, Ann Arbor, MI, USA), Rabbit anti-EP3 antibody, APR-065 (Alomone Labs, Jerusalem, Israel), Rabbit anti-EP3 antibody (PTGER3), NBP1–84835 (Bio-techne, Abingdon, United Kingdom).

Techniques: Real-time Polymerase Chain Reaction

Journal: BMC Cancer

Article Title: Possible predisposition for colorectal carcinogenesis due to altered gene expressions in normal appearing mucosa from patients with colorectal neoplasia

doi: 10.1186/s12885-019-5833-8

Figure Lengend Snippet: Difference in mRNA expression in CRN versus controls

Article Snippet: The following antibodies were tested: Mouse anti-EP3 antibody (5F5), sc-57,105 (Santa Cruz Biotechnology), Rabbit anti-EP3 antibody, 101,760 (Cayman Chemicals, Ann Arbor, MI, USA), Rabbit anti-EP3 antibody, APR-065 (Alomone Labs, Jerusalem, Israel), Rabbit anti-EP3 antibody (PTGER3), NBP1–84835 (Bio-techne, Abingdon, United Kingdom).

Techniques: Expressing