Fig. 3: Low concentrations of Aβo did not affect the total amount of de novo protein synthesis in primary cortical neurons. | Translational Psychiatry

Fig. 3: Low concentrations of Aβo did not affect the total amount of de novo protein synthesis in primary cortical neurons.

From: Low concentrations of amyloid-beta oligomers induce synaptogenesis characteristic for mild cognitive impairment and alter the de novo proteome

Fig. 3: Low concentrations of Aβo did not affect the total amount of de novo protein synthesis in primary cortical neurons.

(a). The strategy for revealing changes in de novo protein synthesis, using the BONCAT method. At DIV 21, primary cortical neurons were daily treated for five days with Aβo (100 nM), or Aβo with eFT508 (2 nM); control cells were not treated. Newly synthesized proteins were labelled by incubation in a methionine-depleted medium containing 1 mM AHA for 4 h prior to harvest. Azide-bearing proteins were subsequently labelled using click chemistry with biotin-alkyne, followed by western blots or proteomic analysis after purification. This figure panel was created using BioRender. (b). Representative BONCAT-Western Blot image showing AHA-labelled newly synthesized proteins across the experimental groups and corresponding total protein levels as revealed by Revert labelling. (c). Quantitative analysis of the results, normalized to total protein levels and presented as bar graphs, indicates no significant changes in the total amount of de novo protein synthesis across the experimental groups. Data represent mean ± SEM, n = 5 or 6 biological replicates. ns = not significant.

Back to article page