Journal: Diabetes

Article Title: Antiatherosclerotic and Renoprotective Effects of Ebselen in the Diabetic Apolipoprotein E/GPx1-Double Knockout Mouse

doi: 10.2337/db10-0195

Figure Lengend Snippet: Ebselen attenuates ApoE −/− GPx1 −/− (dKO) aortic protein levels after 10 weeks of STZ-induced diabetes. Representative immunhistochemical staining for each protein is shown in supplementary online Fig. 2 A–F . Quantitation of immunohistochemical staining within aortic sections is shown for ( A ) Nox2, ( B ) nitrotyrosine (NT), ( C ) 4-HNE, ( D ) VEGF, ( E ) CTGF, and ( F ) VCAM-1. Values are expressed relative to the nondiabetic dKO group which is arbitrarily assigned a value of 1. Bars, mean ± SEM n = 5–7 aortas/group; * P < 0.05 vs. nondiabetic dKO group; # P < 0.05 vs. diabetic dKO group. a.u., arbitrary units; D, diabetic; Eb, ebselen; NT, nitrotyrosine; ND, nondiabetic.

Article Snippet: Nitrotyrosine was assessed in kidney mitochondrial fractions using an ELISA nitrotyrosine immunoassay kit (Oxis Research).

Techniques: Staining, Quantitation Assay, Immunohistochemical staining

Journal: Diabetes

Article Title: Antiatherosclerotic and Renoprotective Effects of Ebselen in the Diabetic Apolipoprotein E/GPx1-Double Knockout Mouse

doi: 10.2337/db10-0195

Figure Lengend Snippet: The H 2 O 2 -mediated increase in nitrotyrosine (NT) is reduced by ebselen in the diabetic ApoE −/− GPx −/− (dKO) kidney. A–D : Representative photomicrographs of kidney tubules after 20 weeks of diabetes. ( A ) nondiabetic dKO, ( B ) diabetic dKO, ( C ) diabetic dKO + Eb, and ( D ) negative control which consisted of species-matched nonimmune IgG in place of primary antibody. E–H : Representative photomicrographs of kidney glomeruli after 20 weeks of diabetes. Nondiabetic dKO ( E ); Diabetic dKO ( F ); diabetic dKO + Eb ( G ); and negative control which consisted of species-matched nonimmune IgG in place of primary antibody ( H ). Quantitation of nitrotyrosine-stained tubules and glomeruli is shown in I and J , respectively. *** P < 0.001; diabetic dKO versus diabetic ApoE −/− kidneys. ## P < 0.01 and ### P < 0.001 vs. diabetic dKO kidneys. Bars, mean ± SEM; n = 6–8 kidneys/group. a.u., arbitrary units; D, diabetic; Eb, ebselen; ND, nondiabetic. (A high-quality digital representation of this figure is available in the online issue.)

Article Snippet: Nitrotyrosine was assessed in kidney mitochondrial fractions using an ELISA nitrotyrosine immunoassay kit (Oxis Research).

Techniques: Negative Control, Quantitation Assay, Staining

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Alpaca ( Vicugna pacos ), the first nonprimate species with a phosphoantigen-reactive Vγ9Vδ2 T cell subset

doi: 10.1073/pnas.1909474117

Figure Lengend Snippet: BTN3-dependent PAg reactivity of transduced alpaca Vγ9Vδ2 TCRs. (A) Murine responder cells (53/4) expressing vpTCR(Jδ2), vpTCR(Jδ4), or a human Vγ9Vδ2 TCR (huTCR) were cocultured with 293T cell lines (WT), 293T BTN3KO, or BTN3KO vpBTN3 (phNGFR mCherry) in the presence of titrated HMBPP doses. Supernatants were tested for mIL-2 after 22 h. (B) In addition, WTH-5 (1 µg/mL) or mIgG2b,κ isotype were added to cultures of TCR transductants with wild-type or BTN3KO vpBTN3 in the presence of 10 µM HMBPP. Duplicates were measured in three experiments and mean + SD are shown. Statistical analysis is shown in SI Appendix, Table S6 for A and a two-way ANOVA (B) was carried out with a Bonferroni post hoc test (ns, not significant: P > 0.05, ****P < 0.0001).

Article Snippet: The activation of TCR transductants was measured via mIL-2 ELISA (BD).

Techniques: Expressing

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Alpaca ( Vicugna pacos ), the first nonprimate species with a phosphoantigen-reactive Vγ9Vδ2 T cell subset

doi: 10.1073/pnas.1909474117

Figure Lengend Snippet: The alpaca B30.2 domain can sense PAgs. (A) A number of 1 × 104 293T BTN3KO or BTN3A1KO cells transduced with huBTN3A1, vpBTN3, hu/vpBTN3, or vp/huBTN3 (all in phNGFR mCherry) were plated overnight. Next, 5 × 104 Murine responder cells [53/4 huTCR or vpTCR(Jδ2)] were added as well as a dilution of HMBPP. Medium was used as a control, and the experiment was carried out in duplicates. Supernatants were tested for mIL-2 after 22 h with mIL-2 ELISA. Means of three independent experiments are shown with SDs. Statistical analysis is summarized in SI Appendix, Table S7. (B) Graphical summary of A with schematic representation of wild-type human (hu) and alpaca BTN3 (vp) and constructed chimeric molecules with an exchange of intracellular domains (hu/vp and vp/hu). The mIL-2 response of responder cells is indicated as high (+++), intermediate (++), low (+), or negative (−).

Article Snippet: The activation of TCR transductants was measured via mIL-2 ELISA (BD).

Techniques: Transduction, Enzyme-linked Immunosorbent Assay, Construct