mouse anti rabbit t lymphocytes fitc 1  (Bio-Rad)

Journal: Cell Reports Medicine

Article Title: Non-overlapping epitopes on the gHgL-gp42 complex for the rational design of a triple-antibody cocktail against EBV infection

doi: 10.1016/j.xcrm.2023.101296

Figure Lengend Snippet: 5E3 and the 5E3+6H2+10E4 cocktail provide potent protection against lethal EBV challenge in humanized mice (A) Experimental timeline for CD34 + hematopoietic stem cell (HSC) engraftment, antibody infusion, EBV challenge, and monitoring for various biological and clinical outcomes. A total of 400 μg of 5E3 (n = 6), 10E4 (n = 6), 5E3+6H2+10E4 cocktail (n = 6), AMMO1 (positive control, n = 5), and VRC01 (negative control, n = 5) was administered to NPI mice via intraperitoneal (i.p.) injection 24 h before intravenous (i.v.) challenge with Akata-EBV (25,000 GRUs). Uninfected control mice (n = 3) did not receive an injection of antibody or virus. Body weight, animal survival, EBV DNA copy number in the peripheral blood, and immunophenotype were recorded weekly for 10 weeks. Humanized mice were euthanized at week 10, and spleens were collected for further analysis. (B–D) Body weight (B), survival (C), and EBV DNA copy number in the peripheral blood samples (D) of mice were monitored weekly. Each line in (D) represents an individual mouse; the limit of detection (10 copies/μL) is indicated by a dashed line. Body weight was converted to percentage. (E–I) The percent changes in lymphocyte proportions of hCD45 + (E), hCD19 + (F), hCD3 + (G), hCD8 + (H), and hCD4 + (I) cells in the peripheral blood were analyzed using flow cytometry. All data are from one experimental replicate and presented as the mean ± SEM. Statistical analyses were performed using one-way ANOVA, comparing each group of data with that of the VRC01 group. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001; ns, not significant. 5E3, 10E4, 5E3+6H2+10E4 combination, AMMO1, VRC01, and uninfected control are colored green, purple, deep pink, gray, brown, and black, respectively. See also Figure S3 .

Article Snippet: MOUSE ANTI RABBIT T LYMPHOCYTES:FITC , Bio-Rad , Cat# MCA800F; RRID: AB_321388.

Techniques: Positive Control, Negative Control, Injection, Control, Virus, Flow Cytometry

Journal: Cell Reports Medicine

Article Title: Non-overlapping epitopes on the gHgL-gp42 complex for the rational design of a triple-antibody cocktail against EBV infection

doi: 10.1016/j.xcrm.2023.101296

Figure Lengend Snippet:

Article Snippet: MOUSE ANTI RABBIT T LYMPHOCYTES:FITC , Bio-Rad , Cat# MCA800F; RRID: AB_321388.

Techniques: Marker, Virus, Recombinant, Adjuvant, Red Blood Cell Lysis, Reverse Transcription, Random Hexamer, Luciferase, Software, Cytometry

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

Article Title: Directionality of nucleocytoplasmic transport of the retroviral gag protein depends on sequential binding of karyopherins and viral RNA

doi: 10.1073/pnas.1000304107

Figure Lengend Snippet: Imp-11 and imp-β drive import of Gag-GFP and interact with Gag in vivo. Coimmunoprecipitation of importins with YFP-NC and MA-GFP (A), Gag-GFP (B), ΔNC.Gag-GFP and ΔMAΔNC.Gag-GFP (C), and ΔMA.Gag-GFP (D). I (input, 3% of total lysate), U (unbound), and B (bound) proteins detected by Western blotting (WB) and antibodies used for immunoprecipitation (IP) are indicated. Molecular weight standards in kilodaltons (kDa) are indicated to the left. The experiments were performed three times, and representative blots are shown. (E) QT6 cells were transiently cotransfected with Gag-GFP derivatives and HA.imp-β or HA.imp-11. HA-tagged importins were detected by rhodamine-conjugated anti-HA, and cells were imaged through the nuclear plane using confocal microscopy. White arrows indicate cells expressing both HA.imp-11 and Gag-GFP, and yellow arrows indicate cells expressing only Gag-GFP. (F) RSV-infected QT6 cells transiently expressed either HA.imp-β or HA.imp-11. RSV Gag was detected with anti-RSV serum and a mouse-anti-rabbit FITC-conjugated secondary antibody. White arrows indicate cells expressing HA-tagged importins and Gag, and yellow arrows point to infected but untransfected cells. The fluorescence intensity of the nucleus was measured in the presence and absence of overexpressed imp-β and imp-11 and divided by the total cellular fluorescence to calculate percent nuclear fluorescence; five cells were measured to calculate the mean.

Article Snippet: Bound proteins were eluted and visualized using either Coomassie blue (Bio-Rad) or Western blotting for H6.Gag.3h utilizing anti-RSV, followed by mouse-anti-rabbit FITC-conjugated secondary antibody.

Techniques: In Vivo, Western Blot, Immunoprecipitation, Molecular Weight, Confocal Microscopy, Expressing, Infection, Fluorescence