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APOE3 and APOE astrocytes were supplemented with 10 µg/mL cholesterol without or with epicatechin (EPI) for 48 hours and cells stained with Hoechst ( A ). Cell clumping ( B ) and cell number ( C ) were measured in n = 3 experiments. D ) APOE astrocytes were treated for 48 hours with 25 nM antimycin A (AA), with and without 25 nM or 2.5 µM epicatechin, or left untreated, and stained with Hoechst and the cell numbers quantified in n = 3 experiments. E ) As D except that <t>methyl-β-cyclodextrin</t> (MβCD) was used in place of epicatechin. F ) APOE3 and APOE astrocytes treated with 5 nM oligomycin (OLI), with and without 5 or 20 µM MβCD, or left untreated for 48 hours and stained with Hoechst, and cell numbers quantified from 3 independent experiments in panel G .
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APOE3 and APOE astrocytes were supplemented with 10 µg/mL cholesterol without or with epicatechin (EPI) for 48 hours and cells stained with Hoechst ( A ). Cell clumping ( B ) and cell number ( C ) were measured in n = 3 experiments. D ) APOE astrocytes were treated for 48 hours with 25 nM antimycin A (AA), with and without 25 nM or 2.5 µM epicatechin, or left untreated, and stained with Hoechst and the cell numbers quantified in n = 3 experiments. E ) As D except that <t>methyl-β-cyclodextrin</t> (MβCD) was used in place of epicatechin. F ) APOE3 and APOE astrocytes treated with 5 nM oligomycin (OLI), with and without 5 or 20 µM MβCD, or left untreated for 48 hours and stained with Hoechst, and cell numbers quantified from 3 independent experiments in panel G .
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APOE3 and APOE astrocytes were supplemented with 10 µg/mL cholesterol without or with epicatechin (EPI) for 48 hours and cells stained with Hoechst ( A ). Cell clumping ( B ) and cell number ( C ) were measured in n = 3 experiments. D ) APOE astrocytes were treated for 48 hours with 25 nM antimycin A (AA), with and without 25 nM or 2.5 µM epicatechin, or left untreated, and stained with Hoechst and the cell numbers quantified in n = 3 experiments. E ) As D except that <t>methyl-β-cyclodextrin</t> (MβCD) was used in place of epicatechin. F ) APOE3 and APOE astrocytes treated with 5 nM oligomycin (OLI), with and without 5 or 20 µM MβCD, or left untreated for 48 hours and stained with Hoechst, and cell numbers quantified from 3 independent experiments in panel G .
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APOE3 and APOE astrocytes were supplemented with 10 µg/mL cholesterol without or with epicatechin (EPI) for 48 hours and cells stained with Hoechst ( A ). Cell clumping ( B ) and cell number ( C ) were measured in n = 3 experiments. D ) APOE astrocytes were treated for 48 hours with 25 nM antimycin A (AA), with and without 25 nM or 2.5 µM epicatechin, or left untreated, and stained with Hoechst and the cell numbers quantified in n = 3 experiments. E ) As D except that <t>methyl-β-cyclodextrin</t> (MβCD) was used in place of epicatechin. F ) APOE3 and APOE astrocytes treated with 5 nM oligomycin (OLI), with and without 5 or 20 µM MβCD, or left untreated for 48 hours and stained with Hoechst, and cell numbers quantified from 3 independent experiments in panel G .
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APOE3 and APOE astrocytes were supplemented with 10 µg/mL cholesterol without or with epicatechin (EPI) for 48 hours and cells stained with Hoechst ( A ). Cell clumping ( B ) and cell number ( C ) were measured in n = 3 experiments. D ) APOE astrocytes were treated for 48 hours with 25 nM antimycin A (AA), with and without 25 nM or 2.5 µM epicatechin, or left untreated, and stained with Hoechst and the cell numbers quantified in n = 3 experiments. E ) As D except that <t>methyl-β-cyclodextrin</t> (MβCD) was used in place of epicatechin. F ) APOE3 and APOE astrocytes treated with 5 nM oligomycin (OLI), with and without 5 or 20 µM MβCD, or left untreated for 48 hours and stained with Hoechst, and cell numbers quantified from 3 independent experiments in panel G .
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APOE3 and APOE astrocytes were supplemented with 10 µg/mL cholesterol without or with epicatechin (EPI) for 48 hours and cells stained with Hoechst ( A ). Cell clumping ( B ) and cell number ( C ) were measured in n = 3 experiments. D ) APOE astrocytes were treated for 48 hours with 25 nM antimycin A (AA), with and without 25 nM or 2.5 µM epicatechin, or left untreated, and stained with Hoechst and the cell numbers quantified in n = 3 experiments. E ) As D except that <t>methyl-β-cyclodextrin</t> (MβCD) was used in place of epicatechin. F ) APOE3 and APOE astrocytes treated with 5 nM oligomycin (OLI), with and without 5 or 20 µM MβCD, or left untreated for 48 hours and stained with Hoechst, and cell numbers quantified from 3 independent experiments in panel G .
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APOE3 and APOE astrocytes were supplemented with 10 µg/mL cholesterol without or with epicatechin (EPI) for 48 hours and cells stained with Hoechst ( A ). Cell clumping ( B ) and cell number ( C ) were measured in n = 3 experiments. D ) APOE astrocytes were treated for 48 hours with 25 nM antimycin A (AA), with and without 25 nM or 2.5 µM epicatechin, or left untreated, and stained with Hoechst and the cell numbers quantified in n = 3 experiments. E ) As D except that <t>methyl-β-cyclodextrin</t> (MβCD) was used in place of epicatechin. F ) APOE3 and APOE astrocytes treated with 5 nM oligomycin (OLI), with and without 5 or 20 µM MβCD, or left untreated for 48 hours and stained with Hoechst, and cell numbers quantified from 3 independent experiments in panel G .
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APOE3 and APOE astrocytes were supplemented with 10 µg/mL cholesterol without or with epicatechin (EPI) for 48 hours and cells stained with Hoechst ( A ). Cell clumping ( B ) and cell number ( C ) were measured in n = 3 experiments. D ) APOE astrocytes were treated for 48 hours with 25 nM antimycin A (AA), with and without 25 nM or 2.5 µM epicatechin, or left untreated, and stained with Hoechst and the cell numbers quantified in n = 3 experiments. E ) As D except that <t>methyl-β-cyclodextrin</t> (MβCD) was used in place of epicatechin. F ) APOE3 and APOE astrocytes treated with 5 nM oligomycin (OLI), with and without 5 or 20 µM MβCD, or left untreated for 48 hours and stained with Hoechst, and cell numbers quantified from 3 independent experiments in panel G .
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APOE3 and APOE astrocytes were supplemented with 10 µg/mL cholesterol without or with epicatechin (EPI) for 48 hours and cells stained with Hoechst ( A ). Cell clumping ( B ) and cell number ( C ) were measured in n = 3 experiments. D ) APOE astrocytes were treated for 48 hours with 25 nM antimycin A (AA), with and without 25 nM or 2.5 µM epicatechin, or left untreated, and stained with Hoechst and the cell numbers quantified in n = 3 experiments. E ) As D except that methyl-β-cyclodextrin (MβCD) was used in place of epicatechin. F ) APOE3 and APOE astrocytes treated with 5 nM oligomycin (OLI), with and without 5 or 20 µM MβCD, or left untreated for 48 hours and stained with Hoechst, and cell numbers quantified from 3 independent experiments in panel G .

Journal: bioRxiv

Article Title: Elevated cholesterol in APOE4 astrocytes drives mitochondrial cristae collapse and ATP synthase dysfunction

doi: 10.64898/2026.08.04.742391

Figure Lengend Snippet: APOE3 and APOE astrocytes were supplemented with 10 µg/mL cholesterol without or with epicatechin (EPI) for 48 hours and cells stained with Hoechst ( A ). Cell clumping ( B ) and cell number ( C ) were measured in n = 3 experiments. D ) APOE astrocytes were treated for 48 hours with 25 nM antimycin A (AA), with and without 25 nM or 2.5 µM epicatechin, or left untreated, and stained with Hoechst and the cell numbers quantified in n = 3 experiments. E ) As D except that methyl-β-cyclodextrin (MβCD) was used in place of epicatechin. F ) APOE3 and APOE astrocytes treated with 5 nM oligomycin (OLI), with and without 5 or 20 µM MβCD, or left untreated for 48 hours and stained with Hoechst, and cell numbers quantified from 3 independent experiments in panel G .

Article Snippet: After 24 h, cells were treated with the following compounds at the indicated final concentrations: cholesterol (10 μg/mL from the Abcam kit; or its equivalent 3 x Gibco cholesterol supplement Cat # 12531018), 25 nM antimycin A (Sigma-Aldrich), 5 nM oligomycin (Sigma-Aldrich), 5 μM or 20 μM β-cyclodextrin (MedChemExpress), and 25 nM or 2.5 μM epicatechin (MedChemExpress).

Techniques: Staining