Journal: Frontiers in Molecular Neuroscience

Article Title: Evaluation of the amyloid beta-GFP fusion protein as a model of amyloid beta peptides-mediated aggregation: a study of DNAJB6 chaperone

doi: 10.3389/fnmol.2015.00040

Figure Lengend Snippet: Canonical members of HSPB family prevent the aggregation of Aβ-GFP. Cells were transfected with HSPB1, HSPB5, HSPB7 or FRTTO at 1:3 ratio for 48 h. Quantification of the pellet/soluble ratio of Aβ-GFP relative to FRTTO was depicted in the chart above the blot. Values represent mean ± SE of two independent experiments. ** P < 0.01, ns = non significant.

Article Snippet: The membranes were blocked with 5% dry milk in PBS with 0.1% Tween 20 (PBST) for 1 h at room temperature and incubated overnight at 4°C with the following primary antibodies: 6E10 (1:1000 in TBST, Covance), anti V5 (1:5000 in PBST, Invitrogen), anti β-actin (1:1000 in PBST, Abcam), anti HSPB1 (1:1000 in PBST, Stress Marq Biosciences), anti HSPB5 (1:1000 in PBST, Stress Marq Biosciences) and anti HSPB7 (1:1000, Abnova).

Techniques: Transfection

Journal: bioRxiv

Article Title: Peripherally expressed misfolded proteins remotely disrupt brain function and aggravate stroke-induced brain injury

doi: 10.1101/785477

Figure Lengend Snippet: a Protein aggregates in primary neuronal cultures detected with a Proteostat aggresome detection kit following incubation with the exosomes isolated from CryAB R120G mice. b Quantitative analysis of a . c Cultured neurons treated with CryAB R120G exosomes showed a reduced ATP level. d Schematic illustration of an cell culture assay to determine the prion-like property of CryAB R120G protein. e Immunocytochemical staining of transfected myc-flag-tagged wild-type mouse CyAB in neural cells following transduction of CryAB R120G aggregate seeds. f Quantitative analysis of e . g Hypothetical model to explain the prion-like propagation of misfolded CryAB R120G protein from the heart to brain. Exosomes containing CryAB R120G protein can be transported from the heart to brain through blood circulation. It is also possible that CryAB R120G protein disrupts BBB and causes CryAB R120G crossing the BBB and entering the brain. In the brain, CryAB R120G protein propagates in a prion-like phenomenon, leading to disruption of BBB, increased neuroinflammation and abnormal brain function. Data are shown as mean ± SD; n = 6 for c and n = 3 for b, f . * p < 0.05, ** p < 0.01.

Article Snippet: Cells were transfected with myc-flag-CryAB plasmid (OriGene, #MR201515) encoding the myc-flag-tagged mouse CryAB protein.

Techniques: Incubation, Isolation, Cell Culture, Staining, Transfection, Transduction