Journal: Journal of Cell Science

Article Title: Cdk5–Foxo3 axis: initially neuroprotective, eventually neurodegenerative in Alzheimer's disease models

doi: 10.1242/jcs.185009

Figure Lengend Snippet: Foxo3 signaling in primary neurons. (A) Primary cortical neurons were treated with glutamate (100 µM) for 6 h in the presence or absence of roscovitine and Cdk5 shRNA, followed by immunostaining. Scale bar: 20 μm. (B) The percentage of cells showing nuclear translocation of Cdk5 and Foxo3. *P<0.01, compared with untreated HT22 cells; #P<0.01, compared with glutamate-treated HT22 cells (Student's t-test). (C) Foxo3 promoter activity was determined as described in the Materials and Methods. *P<0.01, compared with untreated HT22 cells (Student's t-test). (D–G) Bim, FasL, MnSOD and APP promoter activities were measured upon various treatments. *P<0.01, compared with untreated primary cortical neurons (Student's t-test); #P<0.01, compared with glutamate-treated HT22 cells (Student's t-test). (H) Foxo3 levels were analyzed upon glutamate (12 h), roscovitine and Cdk5 shRNA treatments. (I) Bim or MnSOD levels were analyzed upon glutamate, roscovitine, Foxo3 shRNA and Cdk5 shRNA treatments for 12 h. (J) Primary cortical neurons cells were treated with 100 µM glutamate for 18 h. Cdk5 shRNA or Foxo3 shRNA were added 30 h prior to glutamate treatment. Aβ(1–42) released into the medium was measured with an ELISA kit. *P<0.01, compared with untreated neurons (Student's t-test). Graphical results are mean±s.e.m. Each experiment was repeated at least three independent times.

Article Snippet: Both APP-swe HT22 cells and primary cortial neurons were treated with glutamate for 18 h. The released β-amyloid was measured using SensoLyte ® Anti-Humanβ-amyloid (1-42) Quantitative ELISA kit (Anaspec, CA) according to the manufacturer's instructions.

Techniques: shRNA, Immunostaining, Translocation Assay, Activity Assay, Enzyme-linked Immunosorbent Assay

Journal: The Journal of Experimental Medicine

Article Title: Tau deletion promotes brain insulin resistance

doi: 10.1084/jem.20161731

Figure Lengend Snippet: Tau deficiency inhibits an anorexigenic effect of brain insulin administration leading to metabolic disturbances. (A) Cumulative food intake for 24 h measured using metabolic cages in tau KO and littermate controls intracerebroventricularly injected first with vehicle and then 2 µl insulin (5 mg/ml; ***, P < 0.001 vs. WT-PBS; °°°, P < 0.001 vs. tau KO/insulin; two-way ANOVA). (B) Cumulated food intake in WT and tau KO mice 24 h after vehicle or insulin brain injection (same animals as in A; *, P < 0.05; **, P < 0.01 vs. WT-PBS; #, P < 0.05 vs. WT/insulin; °°°, P < 0.001 vs. tau KO/insulin; one-way ANOVA, LSD Fisher’s post-hoc test). (C) 48-h body weight variation after vehicle or insulin brain injection in tau KO and WT littermates (same animals as in A; °°°, P < 0.001 vs. tau KO/insulin; two-way ANOVA). Data in A–C show mean ± SEM from 5–10 mice per group from three independent experiments. (D) Mean food intake (*, P < 0.05, two-way ANOVA). (E) Ambulatory activity (*, P < 0.05, two-way ANOVA). (F) Body weight gain (***, P < 0.05, two-way ANOVA). (G) Plasma leptin levels (**, P < 0.01, Student’s t test). (H and I) Adipose tissue weight (*, P < 0.05; **, P < 0.01, Student’s t test). (J) Glycemia. (K) Insulinemia (*, P < 0.05, Student’s t test). (L) Intraperitoneal glucose tolerance test (***, P < 0.05, two-way ANOVA) in tau KO mice and littermate controls. Quantifications represent mean ± SEM. Controls are indicated as open circles/bars, tau KO as black circles/bars. Dashed lines/bars represent insulin-treated animals. Mice were 6–8 mo old at time of experiments and sacrifice. Metabolic data in C–L show mean ± SEM from 10–12 (D), 9–12 (E), 13–27 (F), 4–5 (G), 11–14 (H and I), 15–17 (J), 20–23 (K), and 12–14 (L) mice per group acquired from three independent experiments.

Article Snippet: Plasma insulin and leptin were measured using ultrasensitive insulin ELISA (Mercodia AB) and a mouse/rat leptin enzyme immunoassay kit (Spibio), respectively.

Techniques: Injection, Activity Assay, Clinical Proteomics