apr 065  (Alomone Labs)

Journal: Frontiers in Pharmacology

Article Title: Prostaglandin E 2 Impairs P2Y 2 /P2Y 4 Receptor Signaling in Cerebellar Astrocytes via EP3 Receptors

doi: 10.3389/fphar.2017.00937

Figure Lengend Snippet: The structure of agonists and antagonists of EP3 receptors.

Article Snippet: Antibodies against P2Y receptors, P2Y 2 and P2Y 4 receptors, and those against the EP3 receptor were from Alomone Labs (Jerusalem BioPark, Israel), while antibodies against GAPDH were purchased from Cell Signaling Technology (Beverly, MA, United States).

Techniques:

Journal: Frontiers in Pharmacology

Article Title: Prostaglandin E 2 Impairs P2Y 2 /P2Y 4 Receptor Signaling in Cerebellar Astrocytes via EP3 Receptors

doi: 10.3389/fphar.2017.00937

Figure Lengend Snippet: Effects of EP3 receptor ligands on UTP calcium responses in rat cerebellar astrocytes. (A) Sulprostone dampened the UTP calcium responses. Astrocytes were stimulated with UTP (100 μM), 30 nM Sulprostone or both these agonists, and the calcium responses were recorded. Where indicated, the cells were pre-incubated with sulprostone for 5 min before UTP stimulation. (B) Effect of L798106, an antagonist of EP3 receptors, on the calcium responses elicited by UTP and on PGE 2 modulation. Where indicated, cells were pre-incubated with L798106 (1 μM) or with L798106 and PGE 2 /sulprostone, prior to stimulation with the nucleotide. Typical calcium traces are shown, and the graphs represent the quantification of the responses obtained as described in Figure and presented as the means ± SEM ( n = 227 cells from three different cultures). ∗∗∗ p < 0.0001 according to one way analysis of variance and Dunnett’s post-test.

Article Snippet: Antibodies against P2Y receptors, P2Y 2 and P2Y 4 receptors, and those against the EP3 receptor were from Alomone Labs (Jerusalem BioPark, Israel), while antibodies against GAPDH were purchased from Cell Signaling Technology (Beverly, MA, United States).

Techniques: Incubation

Journal: Frontiers in Pharmacology

Article Title: Prostaglandin E 2 Impairs P2Y 2 /P2Y 4 Receptor Signaling in Cerebellar Astrocytes via EP3 Receptors

doi: 10.3389/fphar.2017.00937

Figure Lengend Snippet: Prostaglandin E 2 modulates UTP-induced ERK1/2 and PI3-K/Akt activation in astrocytes activated with LPS. Astrocytes were incubated in the presence or absence of LPS (10 ng/ml) for 24 h prior to analyzing the modulatory effect of PGE 2 on the increase in p-ERK1/2 (A) and p-Akt (B) induced by UTP (see Figure ). The Western blots are representative of independent experiments and GAPDH was used as an internal loading control. The values are the means ± SEM from three independent experiments performed on different cultures and the results are expressed relative to the UTP response in each individual experiment. (C) LPS induced COX-2 and EP3 receptor expression in rat cerebellar astrocytes. The presence of COX-2, P2Y2R, P2Y4R, and EP3R proteins was detected in Western blots of total cell lysates. ∗∗∗ p < 0.0001, ∗∗ p < 0.001.

Article Snippet: Antibodies against P2Y receptors, P2Y 2 and P2Y 4 receptors, and those against the EP3 receptor were from Alomone Labs (Jerusalem BioPark, Israel), while antibodies against GAPDH were purchased from Cell Signaling Technology (Beverly, MA, United States).

Techniques: Activation Assay, Incubation, Western Blot, Expressing

Journal: Frontiers in Pharmacology

Article Title: Prostaglandin E 2 Impairs P2Y 2 /P2Y 4 Receptor Signaling in Cerebellar Astrocytes via EP3 Receptors

doi: 10.3389/fphar.2017.00937

Figure Lengend Snippet: Schematic representation of PGE 2 modulation of UTP signaling in rat cerebellar astrocytes. PGE 2 dampened the calcium responses induced by UTP stimulation of P2Y 2 /P2Y 4 , as well as reducing the UTP-induced ERK and Akt activation, and cell migration. The effect of PGE 2 was dependent on EP3 receptor interactions, and it involved EGFR transactivation and PI3K activity, although the involvement of nPKC could be not excluded.

Article Snippet: Antibodies against P2Y receptors, P2Y 2 and P2Y 4 receptors, and those against the EP3 receptor were from Alomone Labs (Jerusalem BioPark, Israel), while antibodies against GAPDH were purchased from Cell Signaling Technology (Beverly, MA, United States).

Techniques: Activation Assay, Migration, Activity Assay