rabbit anti ep3 polyclonal antibody  (Bioss)

Journal: Nature neuroscience

Article Title: NG2 glia protect against prion neurotoxicity by inhibiting microglia-to-neuron prostaglandin E2 signaling.

doi: 10.1038/s41593-024-01663-x

Figure Lengend Snippet: Fig. 7 | PGE2 enhances prion neurotoxicity mainly through the EP4 receptor (Ptger4). a,b, Live-cell imaging (a) and quantitative analysis (b) of chronically prion-infected HovS cells expressing control (Ctrl) transgene or one of the four PGE2 receptors (Ptger1–4). Effects of PGE2 treatment on prion-induced cell toxicity were measured with the ratio of GFP signals under the PGE2 condition against the DMSO condition; n = 4 independent experiments. Data are presented as mean ± s.e.m. One-way ANOVA with Benjamini–Hochberg FDR adjustment for multiple comparisons: P < 0.0001 (Ptger1 versus Ctrl); P = 0.2246 (Ptger2 versus Ctrl); P = 0.3351 (Ptger3 versus Ctrl); P < 0.0001 (Ptger4 versus Ctrl). c, Immunofluorescence of NeuN, Map2 and Tau showing cellular damage of prion- infected primary neurons treated with different concentrations of Ptger4 agonist L902688. d, Quantification of neuronal density as well as Map2-positive and Tau positive areas shown in c; n = 6 independent experiments. Data are presented as

Article Snippet: 3 n atu re p o rtfo lio | rep o rtin g su m m ary A p ril 2 0 2 3 Materials & experimental systems n/a Involved in the study Antibodies Eukaryotic cell lines Palaeontology and archaeology Animals and other organisms Clinical data Dual use research of concern Plants Methods n/a Involved in the study ChIP-seq Flow cytometry MRI-based neuroimaging Antibodies Antibodies used mouse monoclonal antibody against actin (1:10,000, Merck Millipore, MAB1501R, clone C4); mouse monoclonal antibody against PrP (POM1, 1:5000, homemade); rabbit polyclonal antibody against NG2 (1:500, MERCK, AB5320); rabbit polyclonal antibody against PDGFRα (1:500, Santa Cruz, SC-338); rabbit monoclonal antibody against NeuN (1:1000, Abcam, ab177487, clone EPR12763); rabbit polyclonal antibody against NG2 (1:500, a gift from Prof. Stallcup); rabbit polyclonal antibody against Iba1 (1:500, Wako, 019-19741); Rat monoclonal antibody against Cd68 (1:200, BioRad, MCA1957, clone FA-11); rabbit polyclonal antibody against Map2 (1:200, Biolegend, 840601), mouse monoclonal antibody against Cox2 (1:200, Santa Cruz, sc-166475, clone D-12); mouse monoclonal antibody against Ptges (1:200, Santa Cruz, sc-365844, clone H-3); rabbit polyclonal antibody against EP1 (1:200, Bioss Antibodies, BS-6316R); rabbit monoclonal antibody against EP2 (1:200, Abcam, ab167171, clone EPR8030(B)); rabbit polyclonal antibody against EP3 (1:200, Cayman Chemical, 101760); mouse monoclonal antibody against EP4 (1:200, ProteinTech, 66921-1-Ig, clone 4A2A12); rat monoclonal antibody against CD11b antibody (30 ul in 10 ml, ThermoFisher Scientific, 14-0112-82, clone M1/70), mouse monoclonal antibody against Tau (1:200, ThermoFisher Scientific, MN1010, clone BT2), chicken polyclonal antibody against NeuN (1:1000, Merck, ABN91), goat polyclonal antibody against rat IgG (30 ul in 10 m, Jackson ImmunoResearch, 112-005-167); HRP-conjugated goat antirabbit IgG antibody (1:10,000, Jackson ImmunoResearch, 111-035-003); HRP-conjugated goat anti-mouse IgG antibody (1:10,000, Jackson ImmunoResearch, 115-035-003); Alexa488-conjugated goat anti-mouse IgG antibody (1:3000, ThermoFisher Scientific, A32723); Alexa594-conjugated goat anti-rabbit IgG antibody (1:3000, ThermoFisher Scientific, A32740); Alexa647-conjugated goat anti-chicken IgG antibody (1:3000, ThermoFisher Scientific, A32933).

Techniques: Live Cell Imaging, Infection, Expressing, Control, Immunofluorescence

Journal: Journal of Cancer Research and Clinical Oncology

Article Title: Prostaglandin E2 receptor 3 (EP3) signaling promotes migration of cervical cancer via urokinase-type plasminogen activator receptor (uPAR)

doi: 10.1007/s00432-020-03272-0

Figure Lengend Snippet: Effects of EP3 knockdown in HeLa, Siha and C-33A cervical cancer cell lines

Article Snippet: Different primary antibodies were used as follows: rabbit polyclonal anti-EP3 antibody (Abcam, ab94496, 1:500), mouse polyclonal anti-ERK1/2 antibody (Abcam, ab224313, 1:200), rabbit polyclonal anti-p-ERK1/2 antibody (Abcam, ab47339, 1:500), mouse monoclonal anti-p53 antibody (Santa Cruz, OD-1, 1:500) and rabbit polyclonal anti-uPAR antibody (Abcam, ab218106, 1:300). β-actin was used as a housekeeping gene and mouse monoclonal anti-β-actin antibody was diluted as 1:1000 (Sigma, A5441).

Techniques: Knockdown

Journal: Journal of Cancer Research and Clinical Oncology

Article Title: Prostaglandin E2 receptor 3 (EP3) signaling promotes migration of cervical cancer via urokinase-type plasminogen activator receptor (uPAR)

doi: 10.1007/s00432-020-03272-0

Figure Lengend Snippet: EP3 is associated with KEGG signaling pathways of cancer ( a ), calcium signaling ( b ), transforming growth factor-β (TGF-β) ( c ), ECM receptor interaction ( d ), adheren junction ( e ) and cell adhesion molecules (CAMs) ( f ) in carcinogenesis. KEGG pathway gene sets in EP3 high versus low samples were obtained from The Cancer Genome Atlas (TCGA) dataset with the gene set enrichment analysis (GSEA) software ( https://www.software.broadinstitute.org/gsea/index.jsp ). Normalized enrichment score (NES), nominal P value and false discovery rate (FDR) are shown in each plot. The cut-off criteria for GSEA were nominal P value < 0.05 and false discovery rate (FDR) < 0.25

Article Snippet: Different primary antibodies were used as follows: rabbit polyclonal anti-EP3 antibody (Abcam, ab94496, 1:500), mouse polyclonal anti-ERK1/2 antibody (Abcam, ab224313, 1:200), rabbit polyclonal anti-p-ERK1/2 antibody (Abcam, ab47339, 1:500), mouse monoclonal anti-p53 antibody (Santa Cruz, OD-1, 1:500) and rabbit polyclonal anti-uPAR antibody (Abcam, ab218106, 1:300). β-actin was used as a housekeeping gene and mouse monoclonal anti-β-actin antibody was diluted as 1:1000 (Sigma, A5441).

Techniques: Protein-Protein interactions, Software

Journal: Journal of Cancer Research and Clinical Oncology

Article Title: Prostaglandin E2 receptor 3 (EP3) signaling promotes migration of cervical cancer via urokinase-type plasminogen activator receptor (uPAR)

doi: 10.1007/s00432-020-03272-0

Figure Lengend Snippet: EP3 knockdown inhibits the proliferation and migration of cervical cancer cells. a The expression of EP3 is higher in HeLa, SiHa and C-33A than CaSki cells in the protein level by western blots. b The expression of EP3 is higher in HeLa, SiHa and C-33A than CaSki cells in the mRNA level detected by primer I with RT-PCR. c The downregulated expression of EP3 mRNA is shown in HeLa, SiHa and C-33A detected by RT-PCR (* P < 0.05). d BrdU assay suggests the proliferation rate of HeLa and SiHa is decreased by EP3 knockdown compared to the negative control after 48 h. e The proliferation rate of HeLa and SiHa is inhibited followed by stimulation of 100 nM sulprostone and EP3 siRNA compared to the negative control after 48 h (* P < 0.05). f The proliferation rate of SiHa and C-33A is decreased by 100 nM of PGE 2 and L-798,106 compared to the vehicle control after 48 h (0.5% (v/v) DMSO, * P < 0.05). g Representative photographs show the migration of HeLa cells into the wounded area treated with the EP3 siRNA and the negative control after 24 h. h We observed that the relative migration rate of HeLa cells is suppressed in the EP3 siRNA group compared to the negative control (* P < 0.05). i Representative pictures represent the migration of SiHa cells into the wounded area followed by incubating EP3 siRNA and the non-targeting control for 24 h. j The relative migration rate of SiHa cells is inhibited in the EP3 siRNA group compared to the non-targeting control (* P < 0.05). Bar graphs represent mean ± SD ( n = 6). * P < 0.05 is considered as significantly different after comparison between the EP3 siRNA and the negative control (N.C)

Article Snippet: Different primary antibodies were used as follows: rabbit polyclonal anti-EP3 antibody (Abcam, ab94496, 1:500), mouse polyclonal anti-ERK1/2 antibody (Abcam, ab224313, 1:200), rabbit polyclonal anti-p-ERK1/2 antibody (Abcam, ab47339, 1:500), mouse monoclonal anti-p53 antibody (Santa Cruz, OD-1, 1:500) and rabbit polyclonal anti-uPAR antibody (Abcam, ab218106, 1:300). β-actin was used as a housekeeping gene and mouse monoclonal anti-β-actin antibody was diluted as 1:1000 (Sigma, A5441).

Techniques: Knockdown, Migration, Expressing, Western Blot, Reverse Transcription Polymerase Chain Reaction, BrdU Staining, Negative Control, Control, Comparison

Journal: Journal of Cancer Research and Clinical Oncology

Article Title: Prostaglandin E2 receptor 3 (EP3) signaling promotes migration of cervical cancer via urokinase-type plasminogen activator receptor (uPAR)

doi: 10.1007/s00432-020-03272-0

Figure Lengend Snippet: EP3 is correlated with PAI-1 and uPAR in cervical cancer. a, d TIMER database was applied to identify the correlation between EP3 and PAI-1 or uPAR, which is based on the CESC (cervical squamous cell carcinoma and endocervical adenocarcinoma) in the Cancer Genome Atlas (TCGA) dataset ( https://www.cancer.gov ). b, c PAI-1 is associated with poor overall survival (OS) of cervical cancer patients both in GEPIA database ( https://gepia.cancer-pku.cn/ ) and UALCAN database ( https://ualcan.path.uab.edu/index.html ). e, f The association of uPAR with poor prognosis of cervical cancer patients is significant in UALCAN database but not in GEPIA database

Article Snippet: Different primary antibodies were used as follows: rabbit polyclonal anti-EP3 antibody (Abcam, ab94496, 1:500), mouse polyclonal anti-ERK1/2 antibody (Abcam, ab224313, 1:200), rabbit polyclonal anti-p-ERK1/2 antibody (Abcam, ab47339, 1:500), mouse monoclonal anti-p53 antibody (Santa Cruz, OD-1, 1:500) and rabbit polyclonal anti-uPAR antibody (Abcam, ab218106, 1:300). β-actin was used as a housekeeping gene and mouse monoclonal anti-β-actin antibody was diluted as 1:1000 (Sigma, A5441).

Techniques:

Journal: Journal of Cancer Research and Clinical Oncology

Article Title: Prostaglandin E2 receptor 3 (EP3) signaling promotes migration of cervical cancer via urokinase-type plasminogen activator receptor (uPAR)

doi: 10.1007/s00432-020-03272-0

Figure Lengend Snippet: Expression of plasminogen activator inhibitor type 1 (PAI-1) and urokinase-type plasminogen activator receptor (uPAR) is influenced by silencing EP3 gene. a Western blotting analysis shows the expression of phosphorylated extracellular signal-regulated kinases (p-ERK1/2), extracellular signal-regulated kinases (ERK1/2), p53 and uPAR in HeLa and SiHa cells following treatment with EP3 siRNA and the negative control (N.C) for 48 h. β-actin was used as a loading control and all the data was normalized to the β-actin band signals. b PAI-1 levels in the supernatants of HeLa and SiHa cells are enhanced after silencing EP3 compared with the negative control for 48 h by ELISA (* P < 0.05, n = 6). c The histogram illustrates the expression of p-ERK1/2 is increased after silencing EP3 gene for 48 h in SiHa cells (* P < 0.05). d The histogram presents the expression of ERK1/2 is not altered by EP3 siRNA in HeLa and SiHa cells ( P > 0.05). e The histogram illustrates the expression of p53 is inhibited after downregulation of EP3 compared with the negative control for 48 h in SiHa cells (* P < 0.05). f The histogram shows the expression of uPAR is stimulated after EP3 knockdown compared with the negative control for 48 h in SiHa cells (* P < 0.05). Statistically significant differences ( P < 0.05) between EP3 siRNA group and the negative control group are marked with an *. All western blots data are shown as mean ± SD ( n = 3). Full-length blots are shown in Supplementary Fig. 2

Article Snippet: Different primary antibodies were used as follows: rabbit polyclonal anti-EP3 antibody (Abcam, ab94496, 1:500), mouse polyclonal anti-ERK1/2 antibody (Abcam, ab224313, 1:200), rabbit polyclonal anti-p-ERK1/2 antibody (Abcam, ab47339, 1:500), mouse monoclonal anti-p53 antibody (Santa Cruz, OD-1, 1:500) and rabbit polyclonal anti-uPAR antibody (Abcam, ab218106, 1:300). β-actin was used as a housekeeping gene and mouse monoclonal anti-β-actin antibody was diluted as 1:1000 (Sigma, A5441).

Techniques: Expressing, Western Blot, Negative Control, Control, Enzyme-linked Immunosorbent Assay, Knockdown

Journal: Journal of Cancer Research and Clinical Oncology

Article Title: Prostaglandin E2 receptor 3 (EP3) signaling promotes migration of cervical cancer via urokinase-type plasminogen activator receptor (uPAR)

doi: 10.1007/s00432-020-03272-0

Figure Lengend Snippet: Correlation analysis of uPAR and variables

Article Snippet: Different primary antibodies were used as follows: rabbit polyclonal anti-EP3 antibody (Abcam, ab94496, 1:500), mouse polyclonal anti-ERK1/2 antibody (Abcam, ab224313, 1:200), rabbit polyclonal anti-p-ERK1/2 antibody (Abcam, ab47339, 1:500), mouse monoclonal anti-p53 antibody (Santa Cruz, OD-1, 1:500) and rabbit polyclonal anti-uPAR antibody (Abcam, ab218106, 1:300). β-actin was used as a housekeeping gene and mouse monoclonal anti-β-actin antibody was diluted as 1:1000 (Sigma, A5441).

Techniques: Mutagenesis

Journal: Journal of Cancer Research and Clinical Oncology

Article Title: Prostaglandin E2 receptor 3 (EP3) signaling promotes migration of cervical cancer via urokinase-type plasminogen activator receptor (uPAR)

doi: 10.1007/s00432-020-03272-0

Figure Lengend Snippet: Hypothetic schema of EP3 signaling in the migration of human cervical cancer cells. Inhibiting EP3 signaling contributes to phosphorylation of extracellular signal-regulated kinases (p-ERK1/2) and translocation of p53 from the cytoplasm to the nucleus, resulting in an increased transcription of PAI-1. High expression of PAI-1 reduces uPAR cleavage (Magnussen et al. ), thus leading to decreased migration of cervical cancer cells. The EP3 signaling pathway is similar to the one that transforming growth factor-β1 (TGF-β1) induces PAI-1 gene expression via the rapid generation of reactive oxygen species (ROS), phosphorylation of ERK1/2 and the mobilization of p53 signaling (Samarakoon et al. ; Wilkins-Port et al. ). In addition, cytoplasmic p53 is decreased in the cervical cancer cells with high expression of uPAR, which is correlated with poor prognosis in overall survival rates of cervical cancer patients with advanced FIGO stages (III/IV). Therefore, we believed that EP3 signaling regulates the migration of cervical cancer cells through plasminogen activator inhibitor type 1 (PAI-1) and urokinase-type plasminogen activator receptor (uPAR)

Article Snippet: Different primary antibodies were used as follows: rabbit polyclonal anti-EP3 antibody (Abcam, ab94496, 1:500), mouse polyclonal anti-ERK1/2 antibody (Abcam, ab224313, 1:200), rabbit polyclonal anti-p-ERK1/2 antibody (Abcam, ab47339, 1:500), mouse monoclonal anti-p53 antibody (Santa Cruz, OD-1, 1:500) and rabbit polyclonal anti-uPAR antibody (Abcam, ab218106, 1:300). β-actin was used as a housekeeping gene and mouse monoclonal anti-β-actin antibody was diluted as 1:1000 (Sigma, A5441).

Techniques: Migration, Phospho-proteomics, Translocation Assay, Expressing, Gene Expression

Journal: Biochemistry and Biophysics Reports

Article Title: Maternal exposure to prostaglandin E 2 modifies expression of Wnt genes in mouse brain – An autism connection

doi: 10.1016/j.bbrep.2018.03.012

Figure Lengend Snippet: Expression of EP receptors during mouse brain development. RQ values for EP1 , EP2 , EP3α , EP3β , and EP3γ were normalized to RQ values of the EP4 receptor. (A) The EP2 , EP3β , and EP3γ receptors were the most predominantly expressed at Embryonic day 16 (E16). (B-C) Similar pattern was seen at Embryonic day 19 (E19), and Postnatal day 8 (P8). ** p < 0.005, *** p < 0.0005. (A-C) Western blot confirmed protein expression of EP1, EP2, EP3 (all isoforms), and EP4 receptors in the developing brain (right panel). Each value was averaged from three separate experiments (n = 3).

Article Snippet: EP receptors’ protein expression was tested using the primary rabbit polyclonal antibodies anti-EP1 (Santa Cruz Biotechnology), anti-EP2 (Santa Cruz Biotechnology), anti-EP3 (Santa Cruz Biotechnology), and anti-EP4 (Santa Cruz Biotechnology).

Techniques: Expressing, Western Blot