γ secretase inhibitor l 685458  (Tocris)

Journal: Frontiers in bioscience (Landmark edition)

Article Title: The Large Ectodomain of APP Prevents APP from being Directly Cleaved by γ-Secretase.

doi: 10.31083/j.fbl2902078

Figure Lengend Snippet: Fig. 3. The JH is not necessary for the inhibition of γ-cleavage of full-length APP. APPwt (a) and the APPF615P mutant (b) were overexpressed in HEK293 cells and the cells were treated with or without the γ-secretase inhibitor L685458 for 3 h. The cell lysates

Article Snippet: After culturing for 24 h, cells were treated with or without the γ-secretase inhibitor L685458 (MedChemExpress, Monmouth Junction, NY, USA) for 3 h before cell lysis.

Techniques: Inhibition, Mutagenesis

Journal: eLife

Article Title: NHE6 depletion corrects ApoE4-mediated synaptic impairments and reduces amyloid plaque load

doi: 10.7554/eLife.72034

Figure Lengend Snippet:

Article Snippet: Chemical compound, drug , γ-Secretase inhibitor L-685458 , Tocris Bioscience , 2627 , .

Techniques: Staining, Plasmid Preparation, Transfection, Construct, Expressing, shRNA, Recombinant, Sequencing, Software, Imaging

Journal: bioRxiv

Article Title: NHE6-Depletion Corrects ApoE4-Mediated Synaptic Impairments and Reduces Amyloid Plaque Load

doi: 10.1101/2021.03.22.436385

Figure Lengend Snippet: (A,B) Pan-NHE inhibition by EMD87580 or lentiviral knockdown of NHE6 did not alter BACE1 activity in primary neurons of APPswe mice (Tg2576). (A) DIV10 primary neurons were treated with γ-secretase inhibitor L-685458, EMD87580, and/or ApoE4 (as indicated) and harvested for immunoblotting against Aβ-containing C-terminal fragment of APP (βCTF). β-actin was blotted as loading control. Bar graph shows the statistics of n = 3 experiments. (B) Primary neurons of APPswe mice were infected with lentivirus for shRNA expression directed against NHE6 (shNHE6) or a scramble control sequence (-) at DIV7. At DIV13 neurons were treated with L-685458 over night and harvested for immunoblotting against NHE6 and βCTF on DIV14. RAP was blotted as loading control. Bar graph shows the statistics of n = 6 experiments. All data are expressed as mean ± SEM. Statistical analysis was performed using Student t -test. n.s. = not significant.

Article Snippet: For NHE inhibition DIV10 neurons were treated with 5 µg/ml ApoE4, 3 µM EMD87580 (Merck), and/or 1 µM γ-secretase inhibitor L-685458 (Merck) for 5 hours.

Techniques: Inhibition, Activity Assay, Western Blot, Infection, shRNA, Expressing, Sequencing