Journal: Apoptosis

Article Title: Ursolic acid-mediated changes in glycolytic pathway promote cytotoxic autophagy and apoptosis in phenotypically different breast cancer cells

doi: 10.1007/s10495-017-1353-7

Figure Lengend Snippet: UA- and BA-induced DNA damage ( a ) and DNA damage response ( b, c ) in breast cancer cells. a Comet assay. DNA double strand breaks (DSBs) (neutral comet assay) and DNA single strand breaks (SSBs) (alkaline comet assay) are presented. The percentage of tail DNA was used as a parameter of DNA damage. Bars indicate SD, n = 150, * p < 0.05, ** p < 0.01, *** p < 0.001 compared to the control (ANOVA and Dunnett’s a posteriori test). b pATM and pH2AX (γH2AX) (the activation of ATM and H2AX) were measured using Muse™ Cell Analyzer and Muse™ Multi-Color DNA Damage Kit (Merck Millipore). As a positive control for DNA damage, 24 h treatment with 10 µM etoposide was used (C+). c 53BP1 foci formation was revealed using 53BP1 immunostaining and calculated per nucleus. Box and whisker plots are shown, n = 100, *** p < 0.001 compared to the control (ANOVA and Dunnett’s a posteriori test). UA ursolic acid, BA betulinic acid

Article Snippet: Briefly, UA- and BA-treated cells were fixed and incubated with the primary antibody anti-53BP1 (1:200) (Novus Biologicals) and a secondary antibody conjugated to Texas Red (1:1000) (Thermo Fisher Scientific).

Techniques: Single Cell Gel Electrophoresis, Neutral Comet Assay, Alkaline Single Cell Gel Electrophoresis, Activation Assay, Positive Control, Immunostaining, Whisker Assay

Journal: Journal of Veterinary Internal Medicine

Article Title: Investigation of the presence of specific neural antibodies in dogs with epilepsy or dyskinesia using murine and human assays

doi: 10.1111/jvim.16744

Figure Lengend Snippet: Canine anti‐IgLON5 antibodies. Immunofluorescence studies: Diluted serum (1:20) from a dog from the epilepsy/dyskinesia group (Golden Retriever, female, 3 years) is incubated with IgLON5‐transfected human embryonic kidney (HEK) cells. Binding of antibodies to cell surfaces is visualized by two different secondary anti‐canine antibodies coupled with immunofluorescence dyes and subsequently overlaid. The results demonstrate that there are canine anti‐IgLON5 IgG antibodies. Endpoint titration gave a titer of 1:40 (not shown). (A) anti‐canine IgG heavy and light chain (a sensitive secondary antibody), coupled with the red Alexa 594 dye. (B) overlay; yellow: double‐stainings. (C) anti‐canine IgG Fc fragment (the more specific secondary antibody), coupled with the green Alexa 488 dye. Bar: 25 μm.

Article Snippet: Secondary AB was a polyclonal rabbit anti‐dog immunoglobulin‐G (IgG) AB directed against canine IgG heavy and light chains (catalog no. 304‐065‐003; Jackson ImmunoResearch/Dianova) conjugated with red Alexa Fluor 594 used at a dilution of 1:100 and incubated for 30 minutes at room temperature; nuclei were counterstained with Hoechst 33342, 1:10 000.

Techniques: Immunofluorescence, Incubation, Transfection, Binding Assay, Titration

Journal: eLife

Article Title: Atypical memory B-cells are associated with Plasmodium falciparum anemia through anti-phosphatidylserine antibodies

doi: 10.7554/eLife.48309

Figure Lengend Snippet: ( A ) Gating strategy for the characterization of FcRL5 + T-bet + B-cells (CD19 + ) with representative plots of one uninfected control and one P. falciparum patient. ( B ) Percentage of CD19 + FcRL5 + T-bet + B-cells in samples from uninfected controls and P. falciparum patients. Significance assessed by unpaired Student's t test. ****p≤0.0001. 10.7554/eLife.48309.014 Figure 3—source data 1. Source data for .

Article Snippet: Specific B-cell populations were enriched through magnetic bead sorting (Miltenyi) by positive selection with a combination of purified biotinylated anti-FcRL5 antibody (atypical)/anti biotin beads or anti-CD27 (plasma/classical memory cell) coated magnetic beads.

Techniques: Control

Journal: eLife

Article Title: Atypical memory B-cells are associated with Plasmodium falciparum anemia through anti-phosphatidylserine antibodies

doi: 10.7554/eLife.48309

Figure Lengend Snippet: Non-parametric Spearman Correlation analysis of ( A ) atypical MBCs (CD27 – CD21 – FcRL5 + ), ( B ) classical MBCs (CD27 + CD21 + ), ( C ) naïve B-cells (CD27 – CD21 + CD10 – ), ( D ) immature B-cells (CD10 + ), and ( E ) plasma cells or plasmablasts (CD27 + CD21 – CD20 – ) with anti-erythrocyte lysate IgG antibody levels from the plasma of P. falciparum patients. 10.7554/eLife.48309.032 Figure 5—figure supplement 2—source data 1. Source data for .

Article Snippet: Specific B-cell populations were enriched through magnetic bead sorting (Miltenyi) by positive selection with a combination of purified biotinylated anti-FcRL5 antibody (atypical)/anti biotin beads or anti-CD27 (plasma/classical memory cell) coated magnetic beads.

Techniques: Clinical Proteomics

Journal: eLife

Article Title: Atypical memory B-cells are associated with Plasmodium falciparum anemia through anti-phosphatidylserine antibodies

doi: 10.7554/eLife.48309

Figure Lengend Snippet: Non-parametric Spearman correlation of ( A ) atypical MBCs (CD27 – CD21 – FcRL5 + ), ( B ) classical MBCs (CD27 + CD21 + ), ( C ) naïve B-cells (CD27 – CD21 + CD10 – ), ( D ) immature B-cells (CD10 + ), and ( E ) plasma cells or plasmablasts (CD27 + CD21 – CD20 – ) with anti-DNA lysate IgG antibody levels from the plasma of P. falciparum patients. 10.7554/eLife.48309.034 Figure 5—figure supplement 3—source data 1. Source data for .

Article Snippet: Specific B-cell populations were enriched through magnetic bead sorting (Miltenyi) by positive selection with a combination of purified biotinylated anti-FcRL5 antibody (atypical)/anti biotin beads or anti-CD27 (plasma/classical memory cell) coated magnetic beads.

Techniques: Clinical Proteomics

Journal: eLife

Article Title: Atypical memory B-cells are associated with Plasmodium falciparum anemia through anti-phosphatidylserine antibodies

doi: 10.7554/eLife.48309

Figure Lengend Snippet: ( A ) Percentage of T-bet + FcRL5 + B-cells that expanded from the PBMCs of a healthy naïve donor after in-vitro exposure to either uninfected erythrocyte lysate (uLysate) or P. -falciparum -infected erythrocyte lysate (iLysate). ( B ) ELISPOT of enriched populations for either FcRL5 (gray bars) or CD27 (black bars) from PBMCs of healthy naïve US donors after in-vitro exposure to P.-falciparum -infected erythrocyte lysate (iLysate) (N = 3). ASC, antibody-secreting cells. Significance assessed by unpaired Student's t test. **p≤0.01, ***p≤0.001. 10.7554/eLife.48309.041 Figure 7—source data 1. Source data for .

Article Snippet: Specific B-cell populations were enriched through magnetic bead sorting (Miltenyi) by positive selection with a combination of purified biotinylated anti-FcRL5 antibody (atypical)/anti biotin beads or anti-CD27 (plasma/classical memory cell) coated magnetic beads.

Techniques: In Vitro, Infection, Enzyme-linked Immunospot

Journal: eLife

Article Title: Atypical memory B-cells are associated with Plasmodium falciparum anemia through anti-phosphatidylserine antibodies

doi: 10.7554/eLife.48309

Figure Lengend Snippet: Total number of anti-IgM spots of antibody-secreting cells (ASCs) from either CD27 +- or FcRL5 + -enriched PBMC that were stimulated with P.-falciparum- infected erythrocyte lysate. Significant assessed by unpaired Student's t test, ****p<0.0001.

Article Snippet: Specific B-cell populations were enriched through magnetic bead sorting (Miltenyi) by positive selection with a combination of purified biotinylated anti-FcRL5 antibody (atypical)/anti biotin beads or anti-CD27 (plasma/classical memory cell) coated magnetic beads.

Techniques: Infection

Journal: eLife

Article Title: Atypical memory B-cells are associated with Plasmodium falciparum anemia through anti-phosphatidylserine antibodies

doi: 10.7554/eLife.48309

Figure Lengend Snippet:

Article Snippet: Specific B-cell populations were enriched through magnetic bead sorting (Miltenyi) by positive selection with a combination of purified biotinylated anti-FcRL5 antibody (atypical)/anti biotin beads or anti-CD27 (plasma/classical memory cell) coated magnetic beads.

Techniques: Recombinant, Binding Assay, Software, Clinical Proteomics