Journal: The Journal of Neuroscience

Article Title: Fas/Apo [Apoptosis]-1 and Associated Proteins in the Differentiating Cerebral Cortex: Induction of Caspase-Dependent Cell Death and Activation of NF-κB

doi: 10.1523/jneurosci.19-05-01754.1999

Figure Lengend Snippet: Figure 6. The expression of the Fas-associated proteins RIP (A), FADD (B), FLIP (C), and FasL (D) was examined by Western analysis of 50 mg of detergent-soluble protein obtained from P6 cerebral cortex obtained from four separate animals (lanes 1-4). Bands corresponding to RIP (74 kDa) and FasL (37 kDa) were detected in 10 mg of Jurkat cell lysates (a positive control) and in P6 cortex. FLIP expression in P6 cortex was confirmed by the appearance of a similar band just below the 66 kDa molecular weight marker, corresponding to the expected size of ;60 kDa, in protein extract from COS7 cell transfected with the full-length FLIP plasmid (positive control). In contrast, we did not detect any bands corresponding to the expected size for FADD in the cerebral cortex, although a band of the approximate size (24 kDa) was observed in Jurkat cell-derived protein (a positive control). Mkr, Biotinylated marker.

Article Snippet: Binding of the second, primary antibody was visualized by the appropriate secondary antibody, either a biotinylated donkey anti-rabbit antibody (Jackson ImmunoResearch, 1:500) or horse anti-mouse (Vector Laboratories, 1:1000) coupled to either streptavidin-FITC or rhodamineavidin.

Techniques: Expressing, Western Blot, Positive Control, Molecular Weight, Marker, Transfection, Plasmid Preparation, Derivative Assay

Journal: International Journal of Nanomedicine

Article Title: Vaccination with poly(D,L-lactide-co-glycolide) nanoparticles loaded with soluble Leishmania antigens and modified with a TNFα-mimicking peptide or monophosphoryl lipid A confers protection against experimental visceral leishmaniasis

doi: 10.2147/IJN.S141069

Figure Lengend Snippet: Specific anti-sLiAg antibodies raised in vaccinated–infected BALB/c mice. ( A ) Groups of mice (n=5) were vaccinated with PBS, sLiAg + MPLA, PLGA, PLGA-sLiAg, PLGA-sLiAg-MPLA, p8-PLGA, p8-PLGA-sLiAg or p8-PLGA-sLiAg-MPLA three times with 2-week intervals. Two weeks after the second booster vaccination, mice were challenged with 2×10 7 stationary-phase promastigotes of Leishmania infantum . ( B and E ) Two weeks after the last vaccination and ( C , D , F and G ) 2 and 4 months post-challenge, specific anti-sLiAg IgG1 and IgG2a antibodies were determined in mice serum using ELISA. Samples were run in duplicates. ( H ) IgG2a-to-IgG1 ratio. Results are expressed as titer ± SD, that is, the maximum dilution in which sample OD > cut-off OD. Significant differences between mice vaccinated with the multifunctionalized NPs and PBS control group are indicated by asterisks: * P <0.05 or *** P <0.001. Abbreviations: sLiAg, soluble Leishmania infantum antigens; PBS, phosphate-buffered saline; MPLA, monophosphoryl lipid A; PLGA, poly(D,L-lactide-co-glycolide); p8, eight-amino-acid peptide; NPs, nanoparticles; pc, post-challenge; ELISA, enzyme-linked immunosorbent assay; OD, optical density.

Article Snippet: Then, the plates were incubated with biotinylated anti-mouse IgG1 and IgG2a antibodies (AbD Serotec, Oxford, UK) according to the manufacturer’s instructions for 1 h at 37°C and after washing were treated with horse radish peroxidase (HRP) streptavidin solution (AbD Serotec) for 1 h at 37°C.

Techniques: Infection, Enzyme-linked Immunosorbent Assay, Control, Saline