Journal: The American Journal of Pathology

Article Title: Treatment with Anti–Gremlin 1 Antibody Ameliorates Chronic Hypoxia/SU5416–Induced Pulmonary Arterial Hypertension in Mice

doi: 10.1016/j.ajpath.2013.07.017

Figure Lengend Snippet: List of Taqman Probes

Article Snippet: Antibodies used in this study table ft1 table-wrap mode="anchored" t5 caption a7 Gene Assay ID Mouse Id1 Mm00775963_g1 Pai1 Mm00435860_m1 Grem1 Mm00488615_s1 GAPDH Mm99999915_g1 Human BMPR2 Hs00176148_m1 Grem1 Hs01879841_s1 GAPDH 4333764F Open in a separate window List of Taqman Probes

Techniques:

Journal: Scientific reports

Article Title: Serum anti-SERPINE1 antibody as a potential biomarker of acute cerebral infarction.

doi: 10.1038/s41598-021-01176-8

Figure Lengend Snippet: Figure 2. Comparison of serum SERPINE1 antibody levels between the HDs and the ischemic groups. (a) Serum antibody levels against SERPINE1 protein quantified using amplified luminescence proximity homogeneous assay-linked immunosorbent assay (AlphaLISA) and compared between the HDs and the patients in the TIA, aCI, and cCI groups. The Alpha photon counts represent the antibody levels and are shown using a box-whisker plot. Horizontal lines represent medians, and boxes represent the 25th and 75th percentiles. Whiskers represent the 10th and 90th percentiles, and dots represent outliers. Differences were examined using Dunn’s multiple comparison test with type I error adjustment using the Bonferroni procedure. HDs, healthy donors; TIA, transient ischemic attack; aCI, acute cerebral infarction; cCI, chronic cerebral infarction; *p < 0.05; **p < 0.001; n.s., not significant. The antibody level of SERPINE1 was analyzed on the basis of the receiver- operating characteristic (ROC) curve for comparison between TIA (b) and aCI (c). The numbers in the figures indicate the area under the curve (AUC), cutoff values, sensitivity, and specificity.

Article Snippet: The specimens were incubated with a monoclonal anti-human SERPINE1 antibody (Atlas Antibodies, Stockholm, Sweden), which was used as a primary antibody, at a dilution of 1:100 at 4 °C overnight.

Techniques: Comparison, Amplification, Whisker Assay

Journal: Scientific reports

Article Title: Serum anti-SERPINE1 antibody as a potential biomarker of acute cerebral infarction.

doi: 10.1038/s41598-021-01176-8

Figure Lengend Snippet: Figure 1. Western blot analysis. Affinity-purified glutathione-S-transferase (GST; lane 1) and GST-tagged full length SERPINE1 proteins (lane 2) were electrophoresed with sodium dodecyl sulfate–polyacrylamide (11%) gels, followed by staining with Coomassie Brilliant Blue. (a). Western blotting using anti-GST antibody (b) and anti-SERPINE1 antibody (c). Serum samples of patients with acute cerebral infarction (#7074) (d), a transient ischemic attack (#7096) (e), and of a healthy donor (#7021) (f). Electrophoresed molecular weight markers are also shown in (a) (lane M), and the sizes are shown in the left. The asterisk represents degradation products of SERPINE1 after electrophoresis. The number after # is the anonymized patient number and indicates which patient’s serum was used.

Article Snippet: The specimens were incubated with a monoclonal anti-human SERPINE1 antibody (Atlas Antibodies, Stockholm, Sweden), which was used as a primary antibody, at a dilution of 1:100 at 4 °C overnight.

Techniques: Western Blot, Affinity Purification, Staining, Molecular Weight, Electrophoresis

Journal: Scientific reports

Article Title: Serum anti-SERPINE1 antibody as a potential biomarker of acute cerebral infarction.

doi: 10.1038/s41598-021-01176-8

Figure Lengend Snippet: Figure 3. Association between SERPINE1 antibody levels and clinical data. The associations between SERPINE1 antibody levels and clinical parameters such as old age (a); sex (b); hypertension (c), diabetes (d), hyperlipidemia (e), cardiovascular disease (f); body mass index (BMI) (g); history of smoking (h); and alcohol consumption (i) were examined. The SERPINE1 antibody levels determined using AlphaLISA are shown in the box-whisker plots. The p values were calculated using the Kruskal–Wallis test.

Article Snippet: The specimens were incubated with a monoclonal anti-human SERPINE1 antibody (Atlas Antibodies, Stockholm, Sweden), which was used as a primary antibody, at a dilution of 1:100 at 4 °C overnight.

Techniques: Whisker Assay

Journal: Scientific reports

Article Title: Serum anti-SERPINE1 antibody as a potential biomarker of acute cerebral infarction.

doi: 10.1038/s41598-021-01176-8

Figure Lengend Snippet: Figure 4. Immunohistochemical staining for SERPINE1 in an arteriosclerotic plaque. Immunohistochemical staining of atherosclerotic plaques samples from nine patients at the Chiba Cerebral and Cardiovascular Center was performed. A representative case is presented here. These samples were obtained from a different cohort than that from which the sera were collected. Atherosclerotic plaque resected during carotid endarterectomy was immunostained using anti-CD31 (a) or anti-SERPINE1 antibodies (b). Arrows indicate vascular endothelial cells labeled with CD31 antibody, and correspondingly, SERPINE1 was stained by anti-SERPINE1 antibody.

Article Snippet: The specimens were incubated with a monoclonal anti-human SERPINE1 antibody (Atlas Antibodies, Stockholm, Sweden), which was used as a primary antibody, at a dilution of 1:100 at 4 °C overnight.

Techniques: Immunohistochemical staining, Staining, Labeling

Journal: Scientific reports

Article Title: Serum anti-SERPINE1 antibody as a potential biomarker of acute cerebral infarction.

doi: 10.1038/s41598-021-01176-8

Figure Lengend Snippet: Figure 5. Comparison of serum SERPINE1 antigen levels between the aCI and HDs groups. (a) Serum SERPINE1 antigen levels were examined using sandwich enzyme-linked immunosorbent assay in healthy donors and patients with acute cerebral infarction. The results are shown in a box-whisker plot. The p values were calculated using the Kruskal–Wallis test. (b) SERPINE1 antibody levels were plotted against SERPINE1 antigen levels in a scattered plot. The correlation coefficient (r value) and p value were calculated using the Pearson product–moment correlation coefficient.

Article Snippet: The specimens were incubated with a monoclonal anti-human SERPINE1 antibody (Atlas Antibodies, Stockholm, Sweden), which was used as a primary antibody, at a dilution of 1:100 at 4 °C overnight.

Techniques: Comparison, Sandwich ELISA, Whisker Assay

Journal: Oncology Reports

Article Title: Knockdown of SERPINE1 reverses resistance of triple-negative breast cancer to paclitaxel via suppression of VEGFA

doi: 10.3892/or.2020.7770

Figure Lengend Snippet: Identification of signaling pathways and genes related to PTX resistance in breast cancer cell lines. (A) Venn diagram distribution of differentially expressed genes from 2 arrays. As revealed, 518 genes were specifically differentially expressed in GSE28784 cells and 1,778 genes were specifically differentially expressed in GSE90564 cells, while 151 genes were expressed in both datasets. (B) GO annotations and a functional pathway analysis of 151 genes performed using GO-terms and KEGG. (C) Gene-set enrichment analysis of common differentially regulated genes from A. (D) Gene interaction network constructed based on the shared gene list from A. Different colors represent different clusters as calculated using the K-means clustering method. Core genes in the network were identified based on their edge connectivity number. (E) Gene interaction map related to SERPINE1 generated using STRING. PTX, paclitaxel; GO, Gene Ontology; KEGG, Kyoto Encyclopedia of Genes and genomes; SERPINE1 , serine protease inhibitor, clade E member 1; STRING, Search Tool for the Retrieval of Interacting Genes.

Article Snippet: The membranes were blocked for 1 h with 5% skimmed milk at room temperature and incubated with anti-SERPINE1 antibody (cat. no. A00637-1; 1:1,000 dilution; Boster Biological Technology), VEGFA (cat. no. BA0407; 1:500 dilution; Wuhan Boster Biological Technology Co., Ltd.), cleaved caspase-3 (product code ab32042; 1:500 dilution; Abcam), Bax (cat. no. A00183; 1:1,000 dilution; Wuhan Boster Biological Technology Co., Ltd.) and β-actin (product code ab8224; 1:1,000 dilution; Abcam) overnight at 4°C.

Techniques: Protein-Protein interactions, Functional Assay, Construct, Generated, Protease Inhibitor

Journal: Oncology Reports

Article Title: Knockdown of SERPINE1 reverses resistance of triple-negative breast cancer to paclitaxel via suppression of VEGFA

doi: 10.3892/or.2020.7770

Figure Lengend Snippet: SERPINE1 is upregulated in PTX-resistant BC cell lines. (A) Survival curves for MDA-MB-231 PTX-resistant and -sensitive cells treated with various concentrations of PTX. *P<0.05, **P<0.01. (B) Survival curves for MCF-7 PTX-resistant and -sensitive cells treated with various concentrations of PTX. **P<0.01, ***P<0.001. Some points have no error bars because the standard deviation is too small in A and B. (C) SERPINE1 mRNA levels in MDA-MB-231 PTX-resistant vs. -sensitive cells as assessed by RT-qPCR. ***P<0.001. (D) SERPINE1 mRNA levels in MCF-7 PTX-resistant vs. -sensitive cells as assessed by RT-qPCR. ***P<0.001. (E) SERPINE1 protein expression in MDA-MB-231 and MCF-7 PTX-resistant vs. -sensitive cells as measured by western blotting. **P<0.01. SERPINE1 , serine protease inhibitor, clade E member 1; PTX, paclitaxel; BC, breast cancer; RT-qPCR, reverse transcription-quantitative polymerase chain reaction. *P<0.05, **P<0.01, ***P<0.001.

Article Snippet: The membranes were blocked for 1 h with 5% skimmed milk at room temperature and incubated with anti-SERPINE1 antibody (cat. no. A00637-1; 1:1,000 dilution; Boster Biological Technology), VEGFA (cat. no. BA0407; 1:500 dilution; Wuhan Boster Biological Technology Co., Ltd.), cleaved caspase-3 (product code ab32042; 1:500 dilution; Abcam), Bax (cat. no. A00183; 1:1,000 dilution; Wuhan Boster Biological Technology Co., Ltd.) and β-actin (product code ab8224; 1:1,000 dilution; Abcam) overnight at 4°C.

Techniques: Standard Deviation, Quantitative RT-PCR, Expressing, Western Blot, Protease Inhibitor, Reverse Transcription, Real-time Polymerase Chain Reaction

Journal: Oncology Reports

Article Title: Knockdown of SERPINE1 reverses resistance of triple-negative breast cancer to paclitaxel via suppression of VEGFA

doi: 10.3892/or.2020.7770

Figure Lengend Snippet: Knockdown of SERPINE1 suppresses PTX-resistant BC cell survival and promotes apoptosis. (A) Western blots revealing the levels of SERPINE1 protein in knockdown cells. **P<0.01. (B) Survival curves for MDA-MB-231 and MCF-7 PTX-resistant cells with/without SERPINE1 suppression and treated with various concentrations of PTX. *P<0.05, **P<0.01. Some points have no error bars because the standard deviation is too small. (C) Flow cytometric detection of cell apoptosis among BC cells with/without SERPINE1 suppression. *P<0.05, ***P<0.001. SERPINE1 , serine protease inhibitor, clade E member 1; PTX, paclitaxel; BC, breast cancer.

Article Snippet: The membranes were blocked for 1 h with 5% skimmed milk at room temperature and incubated with anti-SERPINE1 antibody (cat. no. A00637-1; 1:1,000 dilution; Boster Biological Technology), VEGFA (cat. no. BA0407; 1:500 dilution; Wuhan Boster Biological Technology Co., Ltd.), cleaved caspase-3 (product code ab32042; 1:500 dilution; Abcam), Bax (cat. no. A00183; 1:1,000 dilution; Wuhan Boster Biological Technology Co., Ltd.) and β-actin (product code ab8224; 1:1,000 dilution; Abcam) overnight at 4°C.

Techniques: Knockdown, Western Blot, Standard Deviation, Protease Inhibitor

Journal: Oncology Reports

Article Title: Knockdown of SERPINE1 reverses resistance of triple-negative breast cancer to paclitaxel via suppression of VEGFA

doi: 10.3892/or.2020.7770

Figure Lengend Snippet: SERPINE1 suppression induces apoptosis in PTX-resistant TNBC cells. (A) Levels of cleaved caspase-3 and Bax proteins in the control and SERPINE1 -knockdown MDA-MB-231 PTX-resistant cells. (B) Levels of cleaved caspase-3 and Bax proteins in the control and SERPINE1 -knockdown MCF-7 PTX-resistant cells. (C) Quantification of cleaved caspase-3 protein levels in the control and SERPINE1 -knockdown PTX-resistant cells. **P<0.01. (D) Quantification of Bax protein levels in the control and SERPINE-knockdown PTX-resistant cells. **P<0.01. SERPINE1 , serine protease inhibitor, clade E member 1; PTX, paclitaxel; TNBC, triple-negative breast cancer.

Article Snippet: The membranes were blocked for 1 h with 5% skimmed milk at room temperature and incubated with anti-SERPINE1 antibody (cat. no. A00637-1; 1:1,000 dilution; Boster Biological Technology), VEGFA (cat. no. BA0407; 1:500 dilution; Wuhan Boster Biological Technology Co., Ltd.), cleaved caspase-3 (product code ab32042; 1:500 dilution; Abcam), Bax (cat. no. A00183; 1:1,000 dilution; Wuhan Boster Biological Technology Co., Ltd.) and β-actin (product code ab8224; 1:1,000 dilution; Abcam) overnight at 4°C.

Techniques: Control, Knockdown, Protease Inhibitor

Journal: Oncology Reports

Article Title: Knockdown of SERPINE1 reverses resistance of triple-negative breast cancer to paclitaxel via suppression of VEGFA

doi: 10.3892/or.2020.7770

Figure Lengend Snippet: Downregulation of SERPINE1 decreases the tumorigenic capacity of PTX-resistant BC cells in vivo . (A) Image revealing the appearance of tumors from mice that were inoculated with MDA-MB-231/PTX cells with or without SERPINE1 knockdown. (B) Quantification of tumor volumes at the end of the in vivo tumorigenic assays. **P<0.01. SERPINE1 , serine protease inhibitor, clade E member 1; PTX, paclitaxel; BC, breast cancer.

Article Snippet: The membranes were blocked for 1 h with 5% skimmed milk at room temperature and incubated with anti-SERPINE1 antibody (cat. no. A00637-1; 1:1,000 dilution; Boster Biological Technology), VEGFA (cat. no. BA0407; 1:500 dilution; Wuhan Boster Biological Technology Co., Ltd.), cleaved caspase-3 (product code ab32042; 1:500 dilution; Abcam), Bax (cat. no. A00183; 1:1,000 dilution; Wuhan Boster Biological Technology Co., Ltd.) and β-actin (product code ab8224; 1:1,000 dilution; Abcam) overnight at 4°C.

Techniques: In Vivo, Knockdown, Protease Inhibitor

Journal: Oncology Reports

Article Title: Knockdown of SERPINE1 reverses resistance of triple-negative breast cancer to paclitaxel via suppression of VEGFA

doi: 10.3892/or.2020.7770

Figure Lengend Snippet: VEGFA is regulated by SERPINE1 and mediates PTX resistance in BC cells. (A) Survival curves for MDA-MB-231 PTX-resistant cells with/without SERPINE1 suppression and supplemented with VEGFA . *P<0.05, **P<0.01. (B) Survival curves for MCF-7/PTX cells with/without SERPINE1 suppression and supplemented with VEGFA . Some points have no error bars because the standard deviation was too small in A and B. *P<0.05, **P<0.01. (C) Flow cytometric evaluation of apoptosis among BC cells with/without SERPINE1 suppression and treated with VEGFA . **P<0.01 vs. NC group; ## P<0.01 vs. sh- SERPINE1 group. (D) Expression of VEGFA protein in BC cells with/without SERPINE1 suppression and treated with VEGFA . **P<0.01. VEGFA , vascular endothelial growth factor A; SERPINE1 , serine protease inhibitor, clade E member 1; PTX, paclitaxel; BC, breast cancer.

Article Snippet: The membranes were blocked for 1 h with 5% skimmed milk at room temperature and incubated with anti-SERPINE1 antibody (cat. no. A00637-1; 1:1,000 dilution; Boster Biological Technology), VEGFA (cat. no. BA0407; 1:500 dilution; Wuhan Boster Biological Technology Co., Ltd.), cleaved caspase-3 (product code ab32042; 1:500 dilution; Abcam), Bax (cat. no. A00183; 1:1,000 dilution; Wuhan Boster Biological Technology Co., Ltd.) and β-actin (product code ab8224; 1:1,000 dilution; Abcam) overnight at 4°C.

Techniques: Standard Deviation, Expressing, Protease Inhibitor