Moreover, cell ethnicities treated with PGE2, LPS or DMSO (S1 Fig) only showed no significant changes in microglial phagocytosis in N9 cells cultured without fA42 treatment. of latex beads ingested and normalized phagocytosis analysis were estimated for each group using FACS analysis. The results are indicated as % LGD-6972 of the untreated control, and are offered as means SEM of three self-employed experiments. Statistical significance was determined by one-way ANOVA followed by Tukeys test.* 0.05 vs the untreated control group; # 0.05 vs the fA42-stimulated group. con, control; PGE2, prostaglandin E2; fA42, fibrillar A peptide (1C42); GW8, GW848687X; AH, AH6809; L-7, L-798106; GW6, GW627368X.(TIF) pone.0147721.s002.tif (728K) GUID:?9FD4EAAC-A787-434A-B36F-4ECD21AE0115 S3 Fig: Dose response curves of agonists of PG receptors EP1-4 in N9 cells. N9 cells were pretreated with dose of agonists of PG receptors EP1-4. Then, cells were stimulated with fA42 (1 M) in the presence or absence of exogenous PGE2 (5 M) for 3 h. Subsequently, cells were subjected to a 1 h process of phagocytosis of fluorescent-labeled latex beads (0.00125%). Average fluorescence intensity of latex beads ingested and normalized phagocytosis analysis were estimated for each group using FACS analysis. The results are indicated as % of the untreated control, and are offered as means SEM of three self-employed experiments. Statistical significance was determined by one-way ANOVA followed by Tukeys test.* 0.05 vs the untreated control group; # 0.05 vs the fA42-stimulated group. con, control; PGE2, prostaglandin E2; fA42, fibrillar A peptide (1C42); PTPE2, 17-phenyl SELP trinor Prostaglandin E2; bu, butaprost; su, sulprostone; L-9, L-902,688.(TIF) pone.0147721.s003.tif (662K) GUID:?C4280238-8BD2-41A0-807F-27F683222691 S4 Fig: Effect of fA42 and curcumin within the production of PGE2 in N9 cells. N9 cells were pretreated with or without curcumin (10 M) for 30 min prior to fA42 (1 M) treatment for 3 h. Enzyme immunoassay of PGE2 was performed as explained in Methods. Experiments were performed with three replicates for each experimental condition. Data are offered relative to control and are offered as means SEM of five self-employed experiments. Statistical significance was determined by two-way ANOVA followed by Tukeys test. con, control; PGE2, prostaglandin E2; fA42, fibrillar A peptide (1C42); Cur, curcumin.(TIF) pone.0147721.s004.tif (327K) GUID:?A09459A0-3CD9-4ECA-A367-8B25C0C3B9DD S5 Fig: Dose response curves of inhibitor and activator of PKA in N9 cells. N9 cells were pretreated with dose of PKA inhibitor H89 or PKA activator 6-Bnz-cAMP for 30 min. Then, cells were stimulated with fA42 (1 M) in the LGD-6972 presence or absence of exogenous PGE2 (5 M) for 3 h. Subsequently, cells were subjected to a 1 h process of phagocytosis of fluorescent-labeled latex beads (0.00125%). The results are LGD-6972 indicated as % of the untreated control, and are offered as means SEM of three self-employed experiments. Statistical significance was determined by one-way ANOVA followed by Tukeys test.* 0.05 vs the untreated control group; # 0.05 vs the fA42-stimulated group. con, control; PGE2, prostaglandin E2; fA42, fibrillar A peptide (1C42); 6-Bnz-cAMP, Adenosine 3?,5?-cyclic Monophosphate, N6-Benzoyl-, Sodium Salt.(TIF) pone.0147721.s005.tif (581K) GUID:?D23B66F8-FC9E-4FA7-AE73-88291FBD7926 Data Availability StatementAll relevant data are within the paper and its Supporting Info files. Abstract Inflammatory activation of microglia and amyloid (A) deposition are considered to work both individually and synergistically to contribute to the improved LGD-6972 risk of Alzheimers disease (AD). Recent studies show that long-term use of phenolic compounds provides safety against AD, primarily because LGD-6972 of the anti-inflammatory actions. We previously suggested that phenolic compound curcumin ameliorated phagocytosis probably through its anti-inflammatory effects rather than direct rules of phagocytic function in electromagnetic field-exposed N9 microglial cells (N9 cells). Here, we explored the prostaglandin-E2 (PGE2)-related signaling pathway that involved in curcumin-mediated phagocytosis in fibrillar -amyloid peptide (1C42) (fA42)-stimulated N9 cells. Treatment with fA42 improved phagocytosis of fluorescent-labeled latex beads in N9 cells. This increase was attenuated inside a dose-dependent manner by endogenous and exogenous PGE2, as well as a selective EP2 or protein kinase A (PKA) agonist, but not by an EP4 agonist. We also found that an antagonist of EP2, but not EP4, abolished the reduction effect of PGE2 on fA42-induced.