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: Frontiers in Oncology

Article Title: ELISA-like QDB method to meet the emerging need of Her2 assessment for breast cancer patients

doi: 10.3389/fonc.2023.920698

Figure Lengend Snippet: Defining the optimized cutoff for 4B5 using receiver operating characteristic (ROC) analysis based on the standard protocol of FISH/IHC analyses. (A) ROC analysis of QDB results. (B) The distribution of HER2 levels within each IHC category. All the FFPE specimens in cohort 1 were categorized as Her2+ and Her2− based on combined IHC and FISH analyses. FISH results superseded the IHC results whenever a conflict occurred. Her2 levels measured by the QDB method using the 4B5 antibody were plotted against categorized Her2 results for ROC analysis ( n = 319) in (A) . In (B) , Her2 levels measured with QDB were plotted against each IHC categorized scores. Log scale was used to illustrate the performance of the proposed cutoff at 0.399 nmol/g. For specimens with undetectable Her2 levels, a value at 0.001 nmol/g was arbitrarily entered to avoid missing any specimen in the graph.

Article Snippet: Rabbit anti-HER2 monoclonal antibody (clone EP3) was purchased from ZSGB-BIO.

Techniques:

Journal: Frontiers in Oncology

Article Title: ELISA-like QDB method to meet the emerging need of Her2 assessment for breast cancer patients

doi: 10.3389/fonc.2023.920698

Figure Lengend Snippet: Absolutely quantitative analysis of Her2 expressions in cohort 2. (A) The distribution of HER2 levels measured with the QDB method using 4B5 and EP3 antibodies respectively in cohort 2. The results were expressed as mean ± SEM. (B) Correlation analysis of the Her2 levels in cohort 2 measured with the QDB method using 4B5 and EP3 antibodies, respectively. Pearson’s correlation analysis was performed with r = 0.981, p < 0.0001. (C) Correlation of HER2 levels measured with QDB using the 4B5 antibody with those from IHC analysis. The results were analyzed using Spearman’s correlation analysis with ρ = 0.545, p < 0.0001.

Article Snippet: Rabbit anti-HER2 monoclonal antibody (clone EP3) was purchased from ZSGB-BIO.

Techniques:

Journal: Frontiers in Oncology

Article Title: ELISA-like QDB method to meet the emerging need of Her2 assessment for breast cancer patients

doi: 10.3389/fonc.2023.920698

Figure Lengend Snippet: Overall distribution of absolutely quantitated HER2 levels in the merged cohort within each IHC category. The Her2 protein levels in both cohort 1 and cohort 2 were measured with the QDB method using EP3 and 4B5 antibodies, respectively, and plotted together against their respective IHC scores. (A) In QDB analysis with the 4B5 antibody, Her2 levels ranged from 0 to 65.05 nmol/g, with a mean of 2.361 ± 0.290 and a median of 0.108, n = 577. (B) For those with the EP3 antibody, its levels ranged from 0 to 31.31 nmol/g, with a mean of 1.419 ± 0.166 and a median of 0.048, n = 578.

Article Snippet: Rabbit anti-HER2 monoclonal antibody (clone EP3) was purchased from ZSGB-BIO.

Techniques:

Journal: Frontiers in Oncology

Article Title: ELISA-like QDB method to meet the emerging need of Her2 assessment for breast cancer patients

doi: 10.3389/fonc.2023.920698

Figure Lengend Snippet: Comparison of dichotomized QDB results measured with 4B5 and EP3 using their respective cutoffs in the merged cohort (cohorts 1 and 2; n = 578).

Article Snippet: Rabbit anti-HER2 monoclonal antibody (clone EP3) was purchased from ZSGB-BIO.

Techniques: Comparison

Journal: Frontiers in Oncology

Article Title: ELISA-like QDB method to meet the emerging need of Her2 assessment for breast cancer patients

doi: 10.3389/fonc.2023.920698

Figure Lengend Snippet: Sensitivity and specificity of Her2 levels from the QDB method using 4B5 and EP3 antibodies, respectively, vs. IHC ( n = 578) using FISH results as the gold standard.

Article Snippet: Rabbit anti-HER2 monoclonal antibody (clone EP3) was purchased from ZSGB-BIO.

Techniques: Immunohistochemistry

Journal: Frontiers in Oncology

Article Title: ELISA-like QDB method to meet the emerging need of Her2 assessment for breast cancer patients

doi: 10.3389/fonc.2023.920698

Figure Lengend Snippet: Evaluation of the distribution of breast cancer specimens with no Her2 expression (Her2-0) from those with any Her2 expression (Her2-E) in each IHC category based on the limit of detection (LOD) of the QDB method using 4B5 antibodies.

Article Snippet: Rabbit anti-HER2 monoclonal antibody (clone EP3) was purchased from ZSGB-BIO.

Techniques: Expressing

Journal: Frontiers in Oncology

Article Title: ELISA-like QDB method to meet the emerging need of Her2 assessment for breast cancer patients

doi: 10.3389/fonc.2023.920698

Figure Lengend Snippet: The distribution of specimens with (Her2-E) and without Her2 expression (Her2-0) stratified by limit of detection (LOD) of QDB analysis using either EP3 or 4B5 antibodies. The LODs of QDB analysis with both 4B5 and EP3 antibodies were at 0.09 nmol/g. Specimens with Her2 levels above 0.09 nmol/g with both antibodies were in gray color while those below 0.09 nmol/g were in the white circle. Specimens with Her2 levels above 0.09 nmol/g only when measured using the 4B5 antibody were in brown color while those only with the EP3 antibody were in blue color.

Article Snippet: Rabbit anti-HER2 monoclonal antibody (clone EP3) was purchased from ZSGB-BIO.

Techniques: Expressing

Journal: Scientific Reports

Article Title: Developing a routine lab test for absolute quantification of HER2 in FFPE breast cancer tissues using Quantitative Dot Blot (QDB) method

doi: 10.1038/s41598-020-69471-4

Figure Lengend Snippet: The clinicopathological characteristics of the patients.

Article Snippet: Rabbit anti-HER2 antibody (clone EP3) was purchased from ZSGB-BIO (Beijing, China).

Techniques:

Journal: Scientific Reports

Article Title: Developing a routine lab test for absolute quantification of HER2 in FFPE breast cancer tissues using Quantitative Dot Blot (QDB) method

doi: 10.1038/s41598-020-69471-4

Figure Lengend Snippet: Correlation of HER2 levels measured with 4B5 and EP3 antibodies. A total of 332 breast cancer FFPE tissues in 2 × 5 µm slices were provided by a local hospital. MCF-7 and BT474 cell lysates were used as internal controls. FFPE tissue lysates (about 0.5 µg/unit) and cell lysates (about 0.3 µg/unit) were applied onto the QDB plates at 2 µl/unit in triplicate for the QDB measurements with clone EP3 and 4B5 respectively. A set of serially diluted HER2 recombinant protein were included in each plate to develop plate-specific standard curve. All results were averaged from three independent experiments, with each sample in triplicate. The correlation of HER2 levels measured with 4B5 and EP3 was analyzed with Pearson’s correlation coefficient analysis using Graphpad software, r = 0.963, p < 0.0001.

Article Snippet: Rabbit anti-HER2 antibody (clone EP3) was purchased from ZSGB-BIO (Beijing, China).

Techniques: Recombinant, Software

Journal: Scientific Reports

Article Title: Developing a routine lab test for absolute quantification of HER2 in FFPE breast cancer tissues using Quantitative Dot Blot (QDB) method

doi: 10.1038/s41598-020-69471-4

Figure Lengend Snippet: Distribution of all 332 samples. HER2 levels in all 332 breast cancer FFPE sample lysates were measured with the QDB method using EP3 antibody. The lysates were diluted to about 0.25 µg/µl, and then 2 µl lysate was used for each sample. ( a ) the distribution of HER2 levels among 332 samples. HER2 levels ranged from 0 (chemiluminescence readings less than two fold over the background) to 31.310 nmol/g. ( b ) All samples were grouped by their IHC scores provided by local hospital. The distributions of HER2 levels in each IHC group were recorded as following: 0, 0–0.205 nmol/g, mean = 0.045 ± 0.006 nmol/g, n = 77; 1 +, 0 –0.410 nmol/g, mean = 0.049 ± 0.008 nmol/g, n = 65; 2+ , 0–7.250 nmol/g, mean = 0.537 ± 0.122 nmol/g, n = 108; and 3+ , 0.329–31.310 nmol/g, mean = 7.120 ± 0.773 nmol/g, n = 82. The intra- and inter-CV were 8.98% and 9.89% respectively.

Article Snippet: Rabbit anti-HER2 antibody (clone EP3) was purchased from ZSGB-BIO (Beijing, China).

Techniques:

Journal: Scientific Reports

Article Title: Developing a routine lab test for absolute quantification of HER2 in FFPE breast cancer tissues using Quantitative Dot Blot (QDB) method

doi: 10.1038/s41598-020-69471-4

Figure Lengend Snippet: Evaluation of QDB results with those of IHC and FISH analyses using ROC analysis. Samples were separated into negative (HER2−) and positive (HER2+) groups based on the recommendations from ASCO/CAP. In ( a ), samples were grouped based on their IHC scores, with 142 samples in the negative group (IHC 0 and 1+), and 82 samples in the positive group (IHC 3+). Absolute HER2 levels from these samples were used for ROC analysis with Graphpad Prism7.0 software. The ROC curve of QDB results was obtained with area under the Curve (AUC) at 0.9998 ± 0.0002; 95% CI 0.9994–1; p < 0.0001. In ( b ), samples were grouped based on both IHC and FISH results, with 214 samples as negative (HER2−) group and 105 samples as positive (HER2+) group. 6 equivocal cases and 7 cases with missing FISH results were excluded in the analysis. Absolute HER2 levels from these samples were used for ROC analysis with Graphpad Prism7.0 software. The area under the curve (AUC) was at 0.9942 ± 0.0031, with 95% CI at 0.9881–1; p < 0.0001. ( c ) The samples were grouped by IHC scores, and the suggested cutoff values from ROC analyses in ( a ) at 0.267 nmol/g (solid line), and in ( b ) at 0.261 nmol/g (dashed line) were shown to demonstrate the effectiveness of these cutoff values to separate samples from HER2+ to HER2− groups. HER2 levels were plotted in log scale to better demonstrate the distribution of QDB results among these samples. For those samples with undetectable HER2 level, a value of 0.001 nmol/g was arbitrarily entered to avoid omitting any sample in the log scale graph.

Article Snippet: Rabbit anti-HER2 antibody (clone EP3) was purchased from ZSGB-BIO (Beijing, China).

Techniques: Software

Journal: Scientific Reports

Article Title: Developing a routine lab test for absolute quantification of HER2 in FFPE breast cancer tissues using Quantitative Dot Blot (QDB) method

doi: 10.1038/s41598-020-69471-4

Figure Lengend Snippet: Assessing HER2 levels by histologic grade. The FFPE specimens (300 out of 332) were grouped according to their Nottingham histologic scores into Grades I, II, and III. The HER2 levels of each grade were used for column statistics analysis with Graphpad Prism7.0 software. The mean ± SD of the HER2 levels were 0.791 ± 0.555 nmol/g for Grade I (n = 36), 1.554 ± 0.330 nmol/g for Grade II (n = 145), and 3.271 ± 0.535 nmol/g for Grade III (n = 119). The statistical difference was assessed with two-tailed Student’s t-test, with p < 0.05 between Grades I and III, and p = 0.005 between Grades II and III. There was no statistical difference between Grade I and Grade II samples.

Article Snippet: Rabbit anti-HER2 antibody (clone EP3) was purchased from ZSGB-BIO (Beijing, China).

Techniques: Software, Two Tailed Test

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