Journal: The American journal of pathology

Article Title: The bradykinin B1 receptor regulates Aβ deposition and neuroinflammation in Tg-SwDI mice.

doi: 10.1016/j.ajpath.2013.01.021

Figure Lengend Snippet: Figure 3 B1R blockade is not associated with changes in APP processing or in levels of LRP1, APOE, and neprilysin in Tg-SwDI mice. Represen- tative blots (A) and quantitative results (B) of Western blot analysis showing that R715 admin- istration (1 mg/kg, s.c., 8 weeks) causes no changes in the levels of APP, APP C-terminal fragments C99 and C83, or APP-cleaving enzymes BACE1, ADAM10, and ADAM17, compared with vehicle-treated animals. C and D: Effect of R715 treatment on the expression of LRP1, APOE, and neprilysin. GAPDH levels were used as loading controls. The values represent means SEM (N Z 6 to 8).

Article Snippet: Western Blot Analysis Equal protein amounts were separated on a 4% to 12% SDSPAGE gradient, transferred to a nitrocellulose membrane, and incubated overnight with primary antibodies at 4 C. The following primary antibodies were used: human APP-CT20, 1741 Passos et al disintegrin and metalloproteinase domainecontaining protein (ADAM)10, ADAM17, b-site APP-cleaving enzyme 1 (BACE1; Calbiochem, San Diego, CA), glyceraldehyde-3phosphate dehydrogenase (GAPDH), B1R (Santa Cruz Biotechnology, Santa Cruz, CA), low-density lipoprotein receptorerelated protein 1 (LRP1), apolipoprotein E (APOE), neprilysin (Abcam, Cambridge, MA), phosphorylated-p65 NF-kB, liver X receptor, peroxisome proliferator-activated receptor g, and matrix metalloproteinase-9 (Cell Signaling Technology, Danvers, MA).

Techniques: Western Blot, Expressing

Journal: Journal of Biological Chemistry

Article Title: ATP-binding Cassette Transporter A1 Mediates the Beneficial Effects of the Liver X Receptor Agonist GW3965 on Object Recognition Memory and Amyloid Burden in Amyloid Precursor Protein/Presenilin 1 Mice*

doi: 10.1074/jbc.m110.108100

Figure Lengend Snippet: FIGURE 2. High dose GW3965 is required for elevated apoE levels in tissue. Cortical and hippocampal extracts from APP/PS1 untreated (: white), APP/PS1 treated (: dark gray), APP/PS1 ABCA1/ untreated (: light gray), and APP/PS1 ABCA1/ treated (: black) mice were immunoblotted for apoE. Left panels are representative blots using undiluted samples from individual mice from the following cohorts: APP/PS1 untreated control: n 7 (cortex), n 8 (hippocampus); APP/PS1 prophylactic: n 8 (cortex) n 8 (hippocam- pus); APP/PS1 low dose therapeutic: n 8 (cortex), n 8 (hippocampus); APP/PS1 high dose therapeutic: n 10 (cortex), n 9 (hippocampus); ABCA1/ APP/PS1 untreated control: n 4 (cortex), n 4 (hippocampus); ABCA1/ APP/PS1 prophylactic: n 8 (cortex) n 8 (hippocampus): ABCA1/ APP/PS1 low dose therapeu- tic: n 7 (cortex), n 6 (hippocampus); ABCA1/ APP/PS1 high dose therapeutic: n 4 (cortex), n 4 (hippocampus). Samples from each mouse were blotted at least twice, normalized to actin, and expressed as fold-difference compared with apoE levels in untreated APP/PS1 controls. Data represent mean S.E., where * represents p 0.05 and ** represents p 0.01 by one-way analysis of variance followed by Tukey post test.

Article Snippet: Western Blot Analysis of ABCA1, ApoE, APP, and APP-CTF— For analysis ofABCA1, apoE, andAPP, 50–100 g of carbonate lysate was electrophoresed through 10% SDS-polyacrylamide gels, transferred to PVDFmembrane (Millipore), and immunodetected using a monoclonal anti-ABCA1 antibody, AC10 (1:1000, a kind gift from Dr. M. R. Hayden), a murine-specific apoE antibody (1:1000, Santa Cruz Biotechnology), an antiAPP antibody that recognizes both murine and human APP (clone 22C11, 1:2000, Chemicon), or an anti-actin antibody (1:5000, Chemicon) as a loading control.

Techniques: Control

Journal: Journal of Biological Chemistry

Article Title: ATP-binding Cassette Transporter A1 Mediates the Beneficial Effects of the Liver X Receptor Agonist GW3965 on Object Recognition Memory and Amyloid Burden in Amyloid Precursor Protein/Presenilin 1 Mice*

doi: 10.1074/jbc.m110.108100

Figure Lengend Snippet: FIGURE 3. ABCA1 is required for increased CSF apoE levels in response to GW3965. Equal volumes of CSF from individual mice from untreated control (C), prophylactic (P), low dose therapeutic (T), and high dose therapeutic (TH) groups of APP/PS1 and APP/PS1 ABCA1/ mice were pooled, separated by 6% native PAGE, and immunoblotted for apoE and albumin. Stokes diameter markers are shown on the right.

Article Snippet: Western Blot Analysis of ABCA1, ApoE, APP, and APP-CTF— For analysis ofABCA1, apoE, andAPP, 50–100 g of carbonate lysate was electrophoresed through 10% SDS-polyacrylamide gels, transferred to PVDFmembrane (Millipore), and immunodetected using a monoclonal anti-ABCA1 antibody, AC10 (1:1000, a kind gift from Dr. M. R. Hayden), a murine-specific apoE antibody (1:1000, Santa Cruz Biotechnology), an antiAPP antibody that recognizes both murine and human APP (clone 22C11, 1:2000, Chemicon), or an anti-actin antibody (1:5000, Chemicon) as a loading control.

Techniques: Control, Clear Native PAGE