Journal: bioRxiv

Article Title: A Highly Immunogenic Measles Virus-based Th1-biased COVID-19 Vaccine

doi: 10.1101/2020.07.11.198291

Figure Lengend Snippet: (A) Schematic depiction of full-length SARS-CoV-2 S and recombinant MeV vac2 genomes used for expression of this antigen (lower schemes). Antigen or antigen encoding genes are depicted in dark grey; MeV viral gene cassettes (in light grey) are annotated. MluI and AatII restriction sites used for cloning of antigen-genes into post P or post H ATU are highlighted (B) Immunoblot analysis of Vero cells infected at an MOI of 0.01 with MeV vac2 -SARS2-S(P), MeV vac2 -SARS2-S(H), or MV vac2 -ATU(P) (MV vac2 ) as depicted above lanes. Uninfected cells served as mock. Blots were probed using rabbit polyclonal anti-SARS spike antibody (upper blot) or mAb reactive against MeV-N (lower blot). Arrows indicate specific bands. ( C, D ) Growth kinetics of recombinant MeV on Vero cells infected at an MOI of 0.03 with MV vac2 -ATU(P) or MeV vac2 -SARS2-S encoding extra genes in post H or post P. Titers of samples prepared at indicated time points post infection were titrated on Vero cells. Means and standard deviations of three to five independent experiments are presented. ( E ) SARS-CoV-2 S protein expression in Vero cells was verified via immunoperoxidase monolayer assay. 50× magnification; scale bar, 500 μm.

Article Snippet: Donkey anti-rabbit IgG-HRP (H&L) polyclonal antibody (1:10,000; 611-7202; Rockland) and goat anti-mouse IgG-HRP (1:10,000; A2554-1ML; Merck, Darmstadt, Germany) served as secondary antibodies.

Techniques: Recombinant, Expressing, Clone Assay, Western Blot, Infection

Journal: Molecular Vision

Article Title: Effect of curcumin on the modulation of αA- and αB-crystallin and heat shock protein 70 in selenium-induced cataractogenesis in Wistar rat pups

doi:

Figure Lengend Snippet: Effects of curcumin on immunohistochemistry of αA- and αB-crystallin in the eye lens of Wistar rat pups exposed to selenium. A : This image shows the control lens incubated in saline alone without any antibody treatment. B : This is a control lens that was incubated with saline and subjected to antibody treatment. C : Lens from rat pups administered with selenium alone. D : Lens from rat pups administered with selenium and curcumin simultaneously. E : Lens from rat pups administered with selenium first and then treated with curcumin after 24 h. F : Lens from rat pups pretreated with curcumin and then administered with selenium after 24 h. Lens sections were preincubated with αA- and αB-crystallin polyclonal antirabbit Immunoglobulin G (IgG) antibody (1:3,000 dilution) and subsequently with goat anti-rabbit IgG-horse radish peroxidase (HRP) conjugate (1:3,000 dilution). The immunoreactivity was developed with 0.01% 3,3-diaminobenzidine tetrahydrochloride (DAB) and H 2 O 2 . Note the brown color formation indicative of peroxidase reaction in the nucleus. Image G , negative control, lens treated with goat anti-rabbit IgG-HRP and developed using DAB and hydrogen peroxide (H 2 O 2 ). The figure shows the high level of αA- and αB-crystallin expression induced by selenium-mediated oxidative stress. This increased crystallin expression and aggregate formation was prevented by curcumin pretreatment.

Article Snippet: After washing with TBS containing 0.05% Tween 20, the sections were incubated with the primary antibody, αA- and αB-crystallin polyclonal anti-rabbit IgG antibody (BD Biosciences, San Jose, CA), at a dilution of 1:500 overnight at 4 °C.

Techniques: Immunohistochemistry, Incubation, Negative Control, Expressing

Journal: Molecular Vision

Article Title: Effect of curcumin on the modulation of αA- and αB-crystallin and heat shock protein 70 in selenium-induced cataractogenesis in Wistar rat pups

doi:

Figure Lengend Snippet: Immunoblot expression of αA- and αB-crystallin in control and experimental group of animals. Lane I, eye lens protein from control (physiologic saline) rat pups (group I); lane II, eye lens protein from selenium-injected rat pups (group II); lane III, eye lens protein from rat pups administered selenium and curcumin simultaneously (group III); lane IV, eye lens protein from rat pups injected with selenium 24 h before being administered with curcumin (group IV); and lane V, eye lens protein from rat pups administered with curcumin 24 h before being injected with selenium (group V). The separated lens protein was preincubated with αA- and αB-crystallin polyclonal antirabbit IgG antibody (1:3,000 dilution) and subsequently with goat antirabbit IgG-HRP (1:3,000 dilution). The immunoreactivity was developed with 0.01% DAB and H 2 O 2 . β-Actin refers to house keeping protein expression and its levels are constant across all treatment groups indicating the normal behaviour of lenses under various treatment. The figure clearly shows increased αA- and αB-crystallin protein expression under selenium-mediated oxidative stress. This increased crystallin protein expression was prevented by curcumin pretreatment.

Article Snippet: After washing with TBS containing 0.05% Tween 20, the sections were incubated with the primary antibody, αA- and αB-crystallin polyclonal anti-rabbit IgG antibody (BD Biosciences, San Jose, CA), at a dilution of 1:500 overnight at 4 °C.

Techniques: Western Blot, Expressing, Injection