Journal: Journal of Atherosclerosis and Thrombosis

Article Title: Phospholipase A 2 Receptor Gene Polymorphisms Alter its Functions and Present a Genetic Risk of an Increased Intima-Media Thickness of the Carotid Artery

doi: 10.5551/jat.34330

Figure Lengend Snippet: Schematic representation of T/C at rs3749117 and G/C at rs35771982 polymorphisms located in the C-type lectin-like domains (CTLD) 1 of PLA 2 R gene

Article Snippet: Human PLA 2 R wild-type plasmid (NM_001195641) was purchased from Origene (Rockville, MD, USA).

Techniques:

Journal: Journal of Atherosclerosis and Thrombosis

Article Title: Phospholipase A 2 Receptor Gene Polymorphisms Alter its Functions and Present a Genetic Risk of an Increased Intima-Media Thickness of the Carotid Artery

doi: 10.5551/jat.34330

Figure Lengend Snippet: Primers used for point mutagenesis of T/C at rs3749117 and G/C at rs35771982 of PLA 2 R gene

Article Snippet: Human PLA 2 R wild-type plasmid (NM_001195641) was purchased from Origene (Rockville, MD, USA).

Techniques: Mutagenesis

Journal: Journal of Atherosclerosis and Thrombosis

Article Title: Phospholipase A 2 Receptor Gene Polymorphisms Alter its Functions and Present a Genetic Risk of an Increased Intima-Media Thickness of the Carotid Artery

doi: 10.5551/jat.34330

Figure Lengend Snippet: Wild-type and mutant human PLA 2 R protein expression in transfected HEK293 cells (A) Western blot analysis of wild-type and mutant PLA 2 R expressed in HEK293 cells using polyclonal antibody for against CTLD5–6 of human PLA 2 R. The apparent molecular weights of wild-type and mutant PLA 2 R are consistent with their calculated molecular weights, indicating efficient translation. No protein band was detected in control mock-transfected cells (mock). (B) Quantitative analysis of the immunoblotting. Protein expression levels of wild-type and mutant PLA 2 R were similar in HEK293 cells transfected with the respective genotypes. (C) Flow cytometric analyses of wild-type and mutant PLA 2 R cell surface expression in HEK293 cells transfected with a cDNA construct encoding wild-type and mutant PLA 2 R using polyclonal antibody against CTLD5–6 of human PLA 2 R. Upper panels, representative dot plots of suspensions of HEK293 cells transfected with wild-type or mutant PLA 2 R; lower panels, representative histograms studying PLA 2 R expression on cells transfected with the respective PLA 2 R genotypes. (D) Comparison of mean fluorescence intensity (MFI) for PLA 2 R expression in HEK293 cells transfected with wild-type or mutant PLA 2 R. (E) PLA 2 R expression in cultures of human umbilical vein endothelial cells (HUVECs), human vascular smooth muscle cells (SMCs), and mock. The mRNA expression levels were quantified with a real-time two-step reverse transcriptase PCR assay. n = 7 in each experiment. NS, statistically not significant.

Article Snippet: Human PLA 2 R wild-type plasmid (NM_001195641) was purchased from Origene (Rockville, MD, USA).

Techniques: Mutagenesis, Expressing, Transfection, Western Blot, Construct, Fluorescence

Journal: Journal of Atherosclerosis and Thrombosis

Article Title: Phospholipase A 2 Receptor Gene Polymorphisms Alter its Functions and Present a Genetic Risk of an Increased Intima-Media Thickness of the Carotid Artery

doi: 10.5551/jat.34330

Figure Lengend Snippet: Comparison of migration, proliferation, and binding to human collagen I between HEK293 cells expressing wild-type PLA 2 R and double mutant PLA 2 R (C at rs3749117 and C at rs35771982) (A) Migration of HEK293 cells expressing wild-type or double mutant type of PLA 2 R on filter membranes coated with PBS (as a control) or collagen I (Coll) in Boyden chamber experiments. (B) Proliferation (measured by light absorbance at 370/492 nm) of cells expressing wild-type or double mutant type of PLA 2 R on dishes coated with PBS or collagen I (Coll). (C) Binding activity to collagen I in cells expressing wild-type or double mutant type of PLA 2 R. Cells were incubated with 125 I-labeled collagen I at 4°C in the absence or presence of a 50-fold excess amount of unlabeled collagen I and washed with cold PBS. The radioactivity of collagen I bound to cells (counts per minute in 100 µ g of protein) is shown after correcting for nonspecific binding. * p < 0.05 in comparison with mock; † p < 0.05 between wild-type; and double mutant type of PLA 2 R. # < 0.05 in comparison with respective genotypes of cells on PBS-coated dishes. n = 6 for each experiment.

Article Snippet: Human PLA 2 R wild-type plasmid (NM_001195641) was purchased from Origene (Rockville, MD, USA).

Techniques: Migration, Binding Assay, Expressing, Mutagenesis, Activity Assay, Incubation, Labeling, Radioactivity

Journal: Journal of Atherosclerosis and Thrombosis

Article Title: Phospholipase A 2 Receptor Gene Polymorphisms Alter its Functions and Present a Genetic Risk of an Increased Intima-Media Thickness of the Carotid Artery

doi: 10.5551/jat.34330

Figure Lengend Snippet: Comparison of migration, proliferation, and binding to human collagen I between HEK293 cells expressing wild-type PLA 2 R and single mutant PLA 2 R (either C at rs3749117 [Singl Mut1] or C at rs35771982 [Singl Mut2]) (A) Migration of HEK293 cells expressing wild-type or single mutant type of PLA 2 R. (B) Proliferation of cells expressing wild-type or single mutant type of PLA 2 R. (C) Binding activity to collagen I in cells expressing wild-type or single mutant type of PLA 2 R. * p < 0.05 in comparison with mock; † p < 0.05 between wild type; and single mutant type of PLA 2 R. # <0.05 in comparison with respective genotypes of cells on PBS-coated dishes. n = 6 for each experiment.

Article Snippet: Human PLA 2 R wild-type plasmid (NM_001195641) was purchased from Origene (Rockville, MD, USA).

Techniques: Migration, Binding Assay, Expressing, Mutagenesis, Activity Assay

Journal: Journal of Atherosclerosis and Thrombosis

Article Title: Phospholipase A 2 Receptor Gene Polymorphisms Alter its Functions and Present a Genetic Risk of an Increased Intima-Media Thickness of the Carotid Artery

doi: 10.5551/jat.34330

Figure Lengend Snippet: Relationships between T/C genotypes at rs3749117 of PLA 2 R gene and an increased maxIMT in male patients

Article Snippet: Human PLA 2 R wild-type plasmid (NM_001195641) was purchased from Origene (Rockville, MD, USA).

Techniques:

Journal: Journal of Atherosclerosis and Thrombosis

Article Title: Phospholipase A 2 Receptor Gene Polymorphisms Alter its Functions and Present a Genetic Risk of an Increased Intima-Media Thickness of the Carotid Artery

doi: 10.5551/jat.34330

Figure Lengend Snippet: Relationships between T/C genotypes at rs3749117 of PLA 2 R gene and an increased maxIMT of the carotid artery in total patient population

Article Snippet: Human PLA 2 R wild-type plasmid (NM_001195641) was purchased from Origene (Rockville, MD, USA).

Techniques:

Journal: Journal of Atherosclerosis and Thrombosis

Article Title: Phospholipase A 2 Receptor Gene Polymorphisms Alter its Functions and Present a Genetic Risk of an Increased Intima-Media Thickness of the Carotid Artery

doi: 10.5551/jat.34330

Figure Lengend Snippet: Relationships between T/C genotypes at rs3749117 of PLA 2 R gene and an increased baPWV (≥ 19 m/sec) in total patient population

Article Snippet: Human PLA 2 R wild-type plasmid (NM_001195641) was purchased from Origene (Rockville, MD, USA).

Techniques:

Journal: Journal of Atherosclerosis and Thrombosis

Article Title: Phospholipase A 2 Receptor Gene Polymorphisms Alter its Functions and Present a Genetic Risk of an Increased Intima-Media Thickness of the Carotid Artery

doi: 10.5551/jat.34330

Figure Lengend Snippet: Relationships between T/C genotypes at rs3749117 of PLA 2 R gene and a decreased ABI ( <0.88) in female patients

Article Snippet: Human PLA 2 R wild-type plasmid (NM_001195641) was purchased from Origene (Rockville, MD, USA).

Techniques:

Journal: Journal of Atherosclerosis and Thrombosis

Article Title: Phospholipase A 2 Receptor Gene Polymorphisms Alter its Functions and Present a Genetic Risk of an Increased Intima-Media Thickness of the Carotid Artery

doi: 10.5551/jat.34330

Figure Lengend Snippet: Univariate and multivariate logistic regression analysis of association of C/C genotype at rs3749117 of PLA 2 R gene and cardiovascular risk factors with an increased maxIMT of the carotid artery in male patients

Article Snippet: Human PLA 2 R wild-type plasmid (NM_001195641) was purchased from Origene (Rockville, MD, USA).

Techniques:

Journal: Journal of Atherosclerosis and Thrombosis

Article Title: Phospholipase A 2 Receptor Gene Polymorphisms Alter its Functions and Present a Genetic Risk of an Increased Intima-Media Thickness of the Carotid Artery

doi: 10.5551/jat.34330

Figure Lengend Snippet: Relationships between T/C genotypes at rs3749117 of PLA 2 R gene and an increased maxIMT in female patients

Article Snippet: Human PLA 2 R wild-type plasmid (NM_001195641) was purchased from Origene (Rockville, MD, USA).

Techniques:

Journal: Journal of Atherosclerosis and Thrombosis

Article Title: Phospholipase A 2 Receptor Gene Polymorphisms Alter its Functions and Present a Genetic Risk of an Increased Intima-Media Thickness of the Carotid Artery

doi: 10.5551/jat.34330

Figure Lengend Snippet: Relationships between T/C genotypes at rs3749117 of PLA 2 R gene and a decreased ABI ( <0.88) in total patient population

Article Snippet: Human PLA 2 R wild-type plasmid (NM_001195641) was purchased from Origene (Rockville, MD, USA).

Techniques:

Journal: Journal of Atherosclerosis and Thrombosis

Article Title: Phospholipase A 2 Receptor Gene Polymorphisms Alter its Functions and Present a Genetic Risk of an Increased Intima-Media Thickness of the Carotid Artery

doi: 10.5551/jat.34330

Figure Lengend Snippet: Relationships between T/C genotypes at rs3749117 of PLA 2 R gene and an increased baPWV (≥ 19 m/sec) in male patients

Article Snippet: Human PLA 2 R wild-type plasmid (NM_001195641) was purchased from Origene (Rockville, MD, USA).

Techniques:

Journal: Journal of Atherosclerosis and Thrombosis

Article Title: Phospholipase A 2 Receptor Gene Polymorphisms Alter its Functions and Present a Genetic Risk of an Increased Intima-Media Thickness of the Carotid Artery

doi: 10.5551/jat.34330

Figure Lengend Snippet: Relationships between T/C genotypes at rs3749117 of PLA 2 R gene and a decreased ABI ( <0.88) in male patients

Article Snippet: Human PLA 2 R wild-type plasmid (NM_001195641) was purchased from Origene (Rockville, MD, USA).

Techniques:

Journal: Journal of Atherosclerosis and Thrombosis

Article Title: Phospholipase A 2 Receptor Gene Polymorphisms Alter its Functions and Present a Genetic Risk of an Increased Intima-Media Thickness of the Carotid Artery

doi: 10.5551/jat.34330

Figure Lengend Snippet: Relationships between T/C genotypes at rs3749117 of PLA 2 R gene and an increased baPWV ( <19.0 m/sec) in female patients

Article Snippet: Human PLA 2 R wild-type plasmid (NM_001195641) was purchased from Origene (Rockville, MD, USA).

Techniques:

Journal: Molecular Cell

Article Title: RIPK1 and Caspase-8 Ensure Chromosome Stability Independently of Their Role in Cell Death and Inflammation

doi: 10.1016/j.molcel.2018.11.010

Figure Lengend Snippet: The Ripoptosome Forms During Normal Mitosis (A–C) Human HT1080 (A), MEFs (B), and HT29 (C) cells were synchronized, and lysates from asynchronous or synchronized cells were immunoprecipitated with anti-Casp8 (HT1080) or anti-FADD (MEFs, HT29) antibodies. Immunoblot analysis using the indicated antibodies is shown. The synchronization scheme and collection points are indicated above. (D) In situ PLA detection of RIPK1 and Casp8 in HT1080 cells. Green dots indicate PLA signals of RIPK1/Casp8 complexes. The panel on the right shows quantifications of RIPK1/Casp8 PLA speckles (mean ± SD from three independent experiments). In each experiment, 10 cells were counted for each mitotic stage. Scale bars: 10 μm. (E–G) In situ PLA detection using antibodies against the indicated proteins. Green dots indicate proximity signals of RIPK1/Casp8 or TRADD/Casp8 in HT1080 (E), RIPK3/FADD, RIPK3/RIPK1, or RIPK3/Casp8 in MEFs (F) or HT29 (G). Scale bars: 10 μm. (H) In situ PLA detection of cFLIP and Casp8 in HT1080 cells. Green dots indicate proximity signals between cFLIP and Casp8. Scale bars: 10 μm. (I) In situ PLA detection of RIPK1 and Casp8 in the indicated cell lines. Green dots indicate PLA signals of RIPK1/Casp8 complexes. Scale bars: 10 μm. (J and K) DEVDase caspase activity analysis using CDK1i-synchronized and released HT1080 cells treated with the indicated conditions.

Article Snippet: Anti-RIPK1 (N-terminal) WB PLA , Cell Signaling , 3493.

Techniques: Immunoprecipitation, Western Blot, In Situ, Activity Assay

Journal: Molecular Cell

Article Title: RIPK1 and Caspase-8 Ensure Chromosome Stability Independently of Their Role in Cell Death and Inflammation

doi: 10.1016/j.molcel.2018.11.010

Figure Lengend Snippet: RIPK1 Interacts with PLK1 (A) HT1080 cells were synchronized with CDK1i and released. Lysates from asynchronous or synchronized HT1080 cells were immunoprecipitated with anti-RIPK1 or anti-PLK1 antibodies. Immunoblot analysis using the indicated antibodies is shown. (B) HT1080 cells were synchronized with CDK1i and released. Lysates from asynchronous or synchronized HT1080 cells were immunoprecipitated with anti-Casp8 antibody. Immunoblot analysis using the indicated antibodies is shown. (C) Lysates from synchronized and released HT29 cells were immunoprecipitated with anti-RIPK1 or anti-FADD antibodies. Immunoblot analysis using the indicated antibodies is shown. (D) The indicated constructs were co-expressed in HEK293T cells. Myc-immunoprecipitation was performed and RIPK1 interaction was assessed via western blot. (E) HT1080 cells were synchronized with CDK1i and released. Lysates from asynchronous or synchronized HT1080 cells were immunoprecipitated with anti-Casp8 antibody. Immunoblot analysis using the indicated antibodies is shown. (F) In situ PLA detection of PLK1/RIPK1 or PLK1/RIPK3 in CDK1i-synchronized and released HT29. Green dots indicate PLA speckles. Scale bars: 10 μm (G) In situ PLA detection of PLK1/RIPK1 or PLK1/RIPK3 in CDK1i-synchronized and released MEFs. Green dots indicate PLA speckles. Scale bars: 10 μm

Article Snippet: Anti-RIPK1 (N-terminal) WB PLA , Cell Signaling , 3493.

Techniques: Immunoprecipitation, Western Blot, Construct, In Situ

Journal: Molecular Cell

Article Title: RIPK1 and Caspase-8 Ensure Chromosome Stability Independently of Their Role in Cell Death and Inflammation

doi: 10.1016/j.molcel.2018.11.010

Figure Lengend Snippet: RIPK1 Negatively Regulates PLK1 (A) Schematic representation of RIPK1- and Casp8-mediated regulation of PLK1. (B) Immunoblots of primary intestinal organoids from two Ripk1 fl/fl,IEC-creERTM animals. Organoids were treated with ETOH (vehicle control) or 4-OHT, synchronized with CDK1i, and released into media. Cells were lysed and analyzed by immunoblotting with the indicated antibodies. (C) In situ PLA detection of PLK1 and RIPK1 in HT1080 cells, following Casp8 or Plk1 siRNA. Green dots indicate PLA signals between PLK1 and RIPK1. Graph shows quantifications of RIPK1/PLK1 PLA speckles (mean ± SD). 15 cells were counted for each condition. Scale bars: 10 μm. (D and E) In vitro cleavage assay. Purified HA-tagged PLK1 construct and recombinant Casp8 were incubated for 1 h. Immunoblots analysis using the indicated antibodies is shown. (F) HT1080 cells were synchronized with CDK1i and released in media containing the indicated drugs. Lysates from asynchronous or synchronized HT1080 cells were immunoprecipitated with anti-Casp8 antibody. Immunoblot analysis using the indicated antibodies is shown. (G) In situ PLA detection of Casp8/RIPK1 in CDK1i-synchronized and released HT1080 cells treated with the indicated agents. Green dots indicate PLA speckles. Scale bars: 10 μm.

Article Snippet: Anti-RIPK1 (N-terminal) WB PLA , Cell Signaling , 3493.

Techniques: Western Blot, In Situ, In Vitro, Cleavage Assay, Purification, Construct, Recombinant, Incubation, Immunoprecipitation

Journal: Molecular Cell

Article Title: RIPK1 and Caspase-8 Ensure Chromosome Stability Independently of Their Role in Cell Death and Inflammation

doi: 10.1016/j.molcel.2018.11.010

Figure Lengend Snippet: RIPK1 Negatively Regulates PLK1-Mediated Phosphorylation of BUBR1 (A) Scheme illustrating how mitotic ripoptosome interacts and modulates PLK1 and how pharmacological inhibition regulates such interaction and downstream substrate activation. Immunofluorescence analysis using anti-BUBR1 or anti-BUBR1-pT680 antibodies. HT1080 cells were synchronized with CDK1i and released into media containing the indicated agents. Scale bars: 10 μm. (B) HT1080 cells were synchronized with CDK1i and released. Lysates from asynchronous or synchronized HT1080 cells were immunoprecipitated with anti-PLK1 antibody. Immunoblot analysis using the indicated antibodies is shown. (C and D) In situ PLA detection of PLK1/BUBR1 (C) or PLK1/BUBR1-pT680 (D) in synchronized HT1080 cells, treated with the indicated agents. Scale bars: 10 μm. (E) Immunofluorescence analysis using anti-BUBR1-pT680 antibodies (under extraction conditions) in CDK1i-synchronized HT1080 (left) and HT29 (right) cells under the indicated RNAi conditions. Cells were released for 30 min, after which MG132 was added for 90 min to arrest cells in metaphase. N.T. indicates non-targeting RNAi Control oligos. Scale bars: 10 μm. (F) Western blot analysis of phosphorylated BUBR1 following knockdown of Control (Ctrl), Ripk1 , or Plk1 in CDK1i-synchronized and released HT1080 cells. (G) CDK1i-synchronized HT1080 cells were released into media containing the indicated agents. Cells were released for 30 min, after which MG132 was added for 90 min to arrest cells in metaphase. Only cells presenting mitotic abnormalities were scored for PLK1 localization. Images show representative examples of PLK1 mis-localization. Scale bars: 10 μm.

Article Snippet: Anti-RIPK1 (N-terminal) WB PLA , Cell Signaling , 3493.

Techniques: Inhibition, Activation Assay, Immunofluorescence, Immunoprecipitation, Western Blot, In Situ

Journal: Molecular Cell

Article Title: RIPK1 and Caspase-8 Ensure Chromosome Stability Independently of Their Role in Cell Death and Inflammation

doi: 10.1016/j.molcel.2018.11.010

Figure Lengend Snippet: Hyper-Activation of RIPK1 Induces Chromosome Mis-Alignment (A) Asynchronized HT1080 cells were pre-incubated for 2 hr with 10 nM SIR-DNA and then treated with the indicated compounds. Mitotic duration of HT1080 assessed by quantifying the time elapsed between nuclear envelope breakdown (NEBD) and anaphase onset following treatment with the indicated agents. (B) Mitotic abnormalities detected in cells imaged by advanced spinning disc confocal microscopy time lapse to determine mitotic timing. Graphs show the percentage of mitotic abnormalities recorded in 100 mitosis per condition. (C) Example of mitotic cells visualized during advance spinning confocal time lapse. Frames were acquired every 6 min. (D) Scheme depicting experimental procedure for synchronization and release of cells during mitosis in indicated drugs. Examples of chromosome alignment defects to illustrate the scoring system. Scale bars: 10 μm. (E and F) CDK1i-synchronized HT1080 (E) and RPE-1 (F) cells were released into media containing the indicated agents. For the analysis in metaphases, cells were released for 30 min, after which MG132 was added for 90 min to arrest cells in metaphase. Anaphases were scored after 2 hr release. Graphs indicate the number (n) of mitosis scored from 3 independent experiments. Statistical analysis was performed via the two-way ANOVA multiple comparison analysis with ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001. Total number of abnormalities was considered in determining statistical significance. Scale bars: 10 μm. (G) Long-term colony formation assay of CDK1i-synchronized HT1080 cells that were released into media containing the indicated drugs for 2 hr. Mitotic cells were collected by shake off, washed, and 1,000 cells were re-plated for clonogenic assay in the absence of drug. Graphs show the mean ± SE of three independent experiments, normalized to control. Two-way ANOVA multiple comparison analysis with ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001.

Article Snippet: Anti-RIPK1 (N-terminal) WB PLA , Cell Signaling , 3493.

Techniques: Activation Assay, Incubation, Confocal Microscopy, Colony Assay, Clonogenic Assay

Journal: Molecular Cell

Article Title: RIPK1 and Caspase-8 Ensure Chromosome Stability Independently of Their Role in Cell Death and Inflammation

doi: 10.1016/j.molcel.2018.11.010

Figure Lengend Snippet: Cells from Ripk1 and Casp8 Knockout Animals Harbor Defects in Chromosome Alignment (A) Grading of segregation defects. (B and C) Chromosome alignment defects of the indicated primary MEFs. Images show representative phenotypes. Cells were released for 30 min after which 10 μM MG132 was added for 90 min to arrest cells in metaphase. Graphs indicate the number (n) of mitosis scored from 3 independent experiments. Statistical analysis was performed via the two-way ANOVA multiple comparison analysis with ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001. Total amount of abnormalities was considered in determining statistical significance Scale bars: 10 μm. (D) Primary intestinal organoids from Ripk1 fl/fl,IEC-creERTM animals. Organoids were treated with ETOH (vehicle control) or 4-OHT, synchronized with CDK1i, released into media, and scored for alignment defects. Cells were released for 30 min, after which 10 μM MG132 was added for 90 min to arrest cells in metaphase. Graphs indicate the number (n) of mitosis scored from 3 independent experiments. Statistical analysis was performed via the two-way ANOVA multiple comparison analysis with ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001. Arrows indicate misaligned chromosomes. Total amount of abnormalities was considered in determining statistical significance. Scale bars: 10 μm. (E) Chromosome alignment defects of the indicated primary MEFs following knockdown of indicated genes. Images show representative phenotypes. Cells were released for 30 min after which 10 μM MG132 was added for 90 min to arrest cells in metaphase. Graphs indicate the number (n) of mitosis scored from 3 independent experiments. Statistical analysis was performed via the two-way ANOVA multiple comparison analysis with ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001. Total amount of abnormalities was considered in determining statistical significance Scale bars: 10 μm. (F) Chromosome alignment defects of the indicated primary MEFs following knockdown of indicated genes. Images show representative phenotypes. Cells were released for 30 min after which 10 μM MG132 was added for 90 min to arrest cells in metaphase. Graphs indicate the number (n) of mitosis scored from 3 independent experiments. Statistical analysis was performed via the two-way ANOVA multiple comparison analysis with ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001. Total amount of abnormalities was considered in determining statistical significance Scale bars: 10 μm. (G–I) Hemotoxylin- and eosin-stained sections of embryos of the indicated age and genotypes. Mitotic abnormalities were scored throughout the entire embryo (G and H) and the large intestine, liver and skin (I). The graph indicates the SE of mitotic abnormalities scored from three (G and H) and two embryos (I). Total amount of abnormalities was considered in determining statistical significance.

Article Snippet: Anti-RIPK1 (N-terminal) WB PLA , Cell Signaling , 3493.

Techniques: Knock-Out, Staining

Journal: Molecular Cell

Article Title: RIPK1 and Caspase-8 Ensure Chromosome Stability Independently of Their Role in Cell Death and Inflammation

doi: 10.1016/j.molcel.2018.11.010

Figure Lengend Snippet: Ripk1 mRNA Levels Correlate with Aneuploidy in Human Cancers (A–D) Bioinformatics analyses of aneuploidy scores in association with RIPK1 mRNA expression (A–B), PLK1 mRNA expression (C), or the normalized ratio of the two (D) in breast (A, C, D) and lung (B) cancer patients. BRCA: breast cancer; LUAD: lung adenocarcinoma. ∗∗ p < 0.01; ∗∗∗ p < 0.001; ∗∗∗∗ p < 0.0001. (A) n : 461; (B) n : 128; (C) n : 293; (D) n : 218.

Article Snippet: Anti-RIPK1 (N-terminal) WB PLA , Cell Signaling , 3493.

Techniques: Expressing

Journal: Molecular Cell

Article Title: RIPK1 and Caspase-8 Ensure Chromosome Stability Independently of Their Role in Cell Death and Inflammation

doi: 10.1016/j.molcel.2018.11.010

Figure Lengend Snippet:

Article Snippet: Anti-RIPK1 (N-terminal) WB PLA , Cell Signaling , 3493.

Techniques: Western Blot, Recombinant, Protease Inhibitor, In Situ, Software

Journal: PLoS ONE

Article Title: An Anti-Phospholipase A 2 Receptor Quantitative Immunoassay and Epitope Analysis in Membranous Nephropathy Reveals Different Antigenic Domains of the Receptor

doi: 10.1371/journal.pone.0061669

Figure Lengend Snippet: Amino Acid Sequences of Consensus PLA 2 R Epitopes Identified by SPOT.

Article Snippet: Western immunoblots were performed on transfected cell lysates (described below) with a commercial goat anti-PLA 2 R (Acris Antibodies, Herford, Germany), mouse anti-GFP (Santa Cruz) and patient sera as described above.

Techniques:

Journal: PLoS ONE

Article Title: An Anti-Phospholipase A 2 Receptor Quantitative Immunoassay and Epitope Analysis in Membranous Nephropathy Reveals Different Antigenic Domains of the Receptor

doi: 10.1371/journal.pone.0061669

Figure Lengend Snippet: Panel A: The median of ALBIA fluorescence units of anti-PLA 2 R positive IMN samples were compared to IMN samples negative for anti-PLA 2 R antibodies and control samples (normal healthy controls and unrelated inflammatory disease controls).Fluorescence values of anti-PLA 2 R positive samples were significantly higher than values of anti-PLA 2 R negative samples and of controls (p-values<0.0001). [Median with interquartile range]. Panel B: ALBIA readings were analyzed according to antibody titres determined on CB-IIF. Samples with a high titre on CB-IIF had often a high fluorescent value on the bead-based assay but ALBIA readings did not correlate with CB-IIF titres. [Whiskers: 2.5–97.5 percentile]. Panel C: A ROC curve is a graphical plot illustrating the performance of a binary classifier system and is used to evaluate diagnostic tests. Sensitivity (fraction of true positives out of positives) is plotted versus 1-specificity (the fraction of false positives out of negatives). Here, our established ALBIA using HEK cell lysates is compared to the EUROIMMUN IIF-CBA. The EUROIMMUNIIF-CBA is a commercially available immunoassay for anti-PLA 2 R and therefore defined the outcome (anti-PLA 2 R positive vs. anti-PLA 2 R negative).Thus, the IIF-CBA perfectly classifies patients with high sensitivity and specificity. With an area under the curve (AUC) of 0.978, the ALBIA is very close to the CB-IIF assay.

Article Snippet: Western immunoblots were performed on transfected cell lysates (described below) with a commercial goat anti-PLA 2 R (Acris Antibodies, Herford, Germany), mouse anti-GFP (Santa Cruz) and patient sera as described above.

Techniques: Fluorescence, Bead-based Assay, Diagnostic Assay

Journal: PLoS ONE

Article Title: An Anti-Phospholipase A 2 Receptor Quantitative Immunoassay and Epitope Analysis in Membranous Nephropathy Reveals Different Antigenic Domains of the Receptor

doi: 10.1371/journal.pone.0061669

Figure Lengend Snippet: Grey scale heat map representation of results from SPOT used to detect PLA 2 R epitopes. Peptide membranes were probed with 10 randomly selected IMN samples that had previously been tested by IIF-CBA for anti-PLA 2 R antibodies (7 positive (IMN+) and 3 negative (IMN-)), as well as 5 normal healthy controls (NHC). The positive control rabbit antibody to PLA 2 R reacted with the expected peptide used as the immunogen. Consensus epitopes and their respective PLA 2 R domains derived from this analysis are illustrated in and summarized in .

Article Snippet: Western immunoblots were performed on transfected cell lysates (described below) with a commercial goat anti-PLA 2 R (Acris Antibodies, Herford, Germany), mouse anti-GFP (Santa Cruz) and patient sera as described above.

Techniques: Positive Control, Derivative Assay

Journal: PLoS ONE

Article Title: An Anti-Phospholipase A 2 Receptor Quantitative Immunoassay and Epitope Analysis in Membranous Nephropathy Reveals Different Antigenic Domains of the Receptor

doi: 10.1371/journal.pone.0061669

Figure Lengend Snippet: All of the determinants identified by epitope mapping were located in the extracellular domain of PLA 2 R and are ∼10 to 25 aa long. Only one epitope is not in the C-type lectin like domains of the receptor. [C-R,cysteine-rich region; FNII, fibronectin type II domain; CTLDs, C-type lectin like domains; N, N-terminal end; C, C-terminal end].

Article Snippet: Western immunoblots were performed on transfected cell lysates (described below) with a commercial goat anti-PLA 2 R (Acris Antibodies, Herford, Germany), mouse anti-GFP (Santa Cruz) and patient sera as described above.

Techniques:

Journal: PLoS ONE

Article Title: An Anti-Phospholipase A 2 Receptor Quantitative Immunoassay and Epitope Analysis in Membranous Nephropathy Reveals Different Antigenic Domains of the Receptor

doi: 10.1371/journal.pone.0061669

Figure Lengend Snippet: Panel A: Different concentrations of a mixture of PLA 2 R derived peptides were used to inhibit the reactivity to the PLA 2 R whole molecule in an addressable laser bead assay. The reactivity showed a significant, dose dependent inhibition. The inhibition with a control peptide (GE-1) was significantly lower. All values are expressed as residual reactivity after inhibition (in %) compared to the sample without inhibitor or control. Panel B: The peptide mixture together with seven individual PLA 2 R peptides and a control peptide were used at a concentration of 126 µg/mL. Besides the peptide mixture, peptide 3 and peptide 4 showed inhibition of anti-PLA 2 R antibodies. All values are expressed as residual reactivity after inhibition (in %) compared to the sample without inhibitor or control. Panel C: The peptide mixture together with seven individual PLA 2 R peptides and a control peptide were used at a concentration of 126 µg/mL. Besides the peptide mixture, peptide 3 and peptide 4 showed inhibition of anti-PLA 2 R antibodies. All values are expressed as ALBIA median fluorescence intensities (MFI) or titer by indirect immunofluorescence on cell-based assay (IIF-CBA).

Article Snippet: Western immunoblots were performed on transfected cell lysates (described below) with a commercial goat anti-PLA 2 R (Acris Antibodies, Herford, Germany), mouse anti-GFP (Santa Cruz) and patient sera as described above.

Techniques: Derivative Assay, Inhibition, Concentration Assay, Fluorescence, Immunofluorescence, Cell Based Assay

Journal: Frontiers in Microbiology

Article Title: Detection of Antilisterial Activity of 3-Phenyllactic Acid Using Listeria innocua as a Model

doi: 10.3389/fmicb.2018.01373

Figure Lengend Snippet: Minimum inhibitory concentration (MIC, mM) of total (PLA tot ) and undissociated (PLAH) phenyllactic acid against Listeria innocua ATCC 33090 detected in BHI at different pH values.

Article Snippet: Minimum inhibitory concentration (MIC) and MBC of PLA against L. innocua ATCC 33090 were evaluated.

Techniques: Concentration Assay

Journal: Frontiers in Microbiology

Article Title: Detection of Antilisterial Activity of 3-Phenyllactic Acid Using Listeria innocua as a Model

doi: 10.3389/fmicb.2018.01373

Figure Lengend Snippet: MIC and MBC values (mM) of total (Ac tot ) and undissociated (AcH) phenyllactic acid (PLA), gallic acid (GA), caffeic acid (CA), ferulic acid (FA), and lactic acid (LA) against L. innocua ATCC 33090 grown in BHI broth at pH 5.5.

Article Snippet: Minimum inhibitory concentration (MIC) and MBC of PLA against L. innocua ATCC 33090 were evaluated.

Techniques:

Journal: Frontiers in Microbiology

Article Title: Detection of Antilisterial Activity of 3-Phenyllactic Acid Using Listeria innocua as a Model

doi: 10.3389/fmicb.2018.01373

Figure Lengend Snippet: Survival of L. innocua ATCC 33090 in (∘) MES buffer (C), or in MES buffer containing: (♢), phenyllactic acid at MIC concentration detected at pH 5.5 (PLA_MIC); (□), lactic acid (LA) at MIC concentration detected at pH 5.5 (LA_MIC) or ( ), LA at the same concentration of MIC of PLA (LA_PLA). Symbols represent the mean values with standard deviation of three independent experiments and the curves represent the survival models obtained with GInaFiT software.

Article Snippet: Minimum inhibitory concentration (MIC) and MBC of PLA against L. innocua ATCC 33090 were evaluated.

Techniques: Concentration Assay, Standard Deviation, Software

Journal: Frontiers in Microbiology

Article Title: Detection of Antilisterial Activity of 3-Phenyllactic Acid Using Listeria innocua as a Model

doi: 10.3389/fmicb.2018.01373

Figure Lengend Snippet: Survival of L. innocua ATCC 33090 in (∘) MSE buffer (C), or in MSE buffer containing: (□), gallic acid (GA_MIC); (△), ferulic acid (FA_MIC); (▽), caffeic acid (CA_MIC) or (♢), phenyllactic acid (PLA_MIC) at MIC concentration detected at pH 5.5. Symbols represent the mean values with standard deviation of three independent experiments and the curves represent the survival models obtained with GInaFiT software.

Article Snippet: Minimum inhibitory concentration (MIC) and MBC of PLA against L. innocua ATCC 33090 were evaluated.

Techniques: Concentration Assay, Standard Deviation, Software

Journal: Frontiers in Microbiology

Article Title: Detection of Antilisterial Activity of 3-Phenyllactic Acid Using Listeria innocua as a Model

doi: 10.3389/fmicb.2018.01373

Figure Lengend Snippet: Survival of L. innocua ATCC 33090 in MES buffer containing: (□), gallic acid (GA_MBC); (△), ferulic acid (FA_MBC); (▽), caffeic acid (CA_MBC) or (♢), phenyllactic acid (PLA_MBC) at MBC concentration detected at pH 5.5. Symbols represent the mean values with standard deviation of three independent experiments and the curves represent the survival models obtained with GInaFiT software.

Article Snippet: Minimum inhibitory concentration (MIC) and MBC of PLA against L. innocua ATCC 33090 were evaluated.

Techniques: Concentration Assay, Standard Deviation, Software

Journal: Frontiers in Microbiology

Article Title: Detection of Antilisterial Activity of 3-Phenyllactic Acid Using Listeria innocua as a Model

doi: 10.3389/fmicb.2018.01373

Figure Lengend Snippet: Zeta potential of L. innocua ATCC 33090 in (∘) ultrapure water (C), or in ultrapure water containing: (□), gallic acid (GA_MBC); (△), ferulic acid (FA_MBC); (▽), caffeic acid (CA_MBC) or (♢), phenyllactic acid (PLA_MBC) at MBC concentration detected at pH 5.5.

Article Snippet: Minimum inhibitory concentration (MIC) and MBC of PLA against L. innocua ATCC 33090 were evaluated.

Techniques: Concentration Assay

Journal: Frontiers in Microbiology

Article Title: Detection of Antilisterial Activity of 3-Phenyllactic Acid Using Listeria innocua as a Model

doi: 10.3389/fmicb.2018.01373

Figure Lengend Snippet: Cellular loss content of L. innocua ATCC 33090 in (∘) ultrapure water (C), or in ultrapure water containing: (□), gallic acid (GA_MBC); (△), ferulic acid (FA_MBC); (▽), caffeic acid (CA_MBC) or (♢), phenyllactic acid (PLA_MBC) at MBC concentration detected at pH 5.5.

Article Snippet: Minimum inhibitory concentration (MIC) and MBC of PLA against L. innocua ATCC 33090 were evaluated.

Techniques: Concentration Assay