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. 2022 Jun 29;14:88. doi: 10.1186/s13195-022-01020-9

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© The Author(s) 2022

Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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Fig. 3 — Small molecule compounds inhibit apoE4-catalyzed Aβ pathology in primary neurons from 5xFAD transgenic mice. a Schematic of drug efficacy experiments using primary neurons from the 5xFAD transgenic mouse model of Alzheimer’s disease. One week after cell isolation from day P1-P2 pups, cells were exposed to 100 nM Aβ42 and 1 nM apoE4 and were treated concurrently with 0.01, 0.1, or 1.0 μM compound in a final concentration of 0.5% (v/v) DMSO. The cell medium was changed every 3 days by removing half and replacing it with a fresh medium containing Aβ42, apoE4, and compound such that the starting concentrations were maintained for the duration of the experiment. At 9 dpe, cells were fixed for immunocytochemistry (ICC) and the conditioned medium was collected for analysis of Aβ concentrations by enzyme-linked immunosorbent assay (ELISA). b Representative ICC images of neurons at 9 dpe labeled for total tau (red), Aβ (white), and cell nuclei (blue). Scale bars = 20 μm. c Percent positive area of Hoechst⁺ cell nuclei at 9 dpe, relative to the DPBS control group. The data represent the mean ± SD of n = 6 wells per group. *P < 0.05 by one-way ANOVA. d Percent positive area of Aβ⁺ pathology at 9 dpe, relative to the DPBS control group. The data represent the mean ± SD of n = 6 wells per group. ***P < 0.001 by one-way ANOVA. e Representative ICC images of neurons at 9 dpe to apoE4 and Aβ42 and treated with compounds at 1 μM and labeled for total tau (red), Aβ (white), and cell nuclei (blue). Scale bars = 20 μm. f Percent positive area of Hoechst⁺ cell nuclei at 9 dpe, relative to the DMSO control group. The data represent the mean ± SD of n = 6 wells for DMSO and n = 3 wells per concentration for compounds. *P < 0.05, **P < 0.01, and ***P < 0.001 compared to the DMSO control by one-way ANOVA. g Percent positive area of Aβ⁺ pathology at 9 dpe, relative to the DMSO control group. The data represent the mean ± SD of n = 6 wells for DMSO and n = 3 wells per concentration for compounds. *P < 0.05, **P < 0.01, and ***P < 0.001 compared to the DMSO control by one-way ANOVA