Three flavones, 7,8-DHF, quercetin (3,3’,4’,5,7-pentahydroxyflavone) and apigenin (4’,5,7-trihydroxyflavone) (Sigma-Aldrich, St. Louis, MO, USA), were tested. The three flavones demonstrated solubility up to 100 μM in cell culture medium. A single copy of the Aβ-green fluorescent protein (GFP) gene was integrated into SH-SY5Y cells (ATCC No. CRL-2266) to create human Aβ folding reporter SH-SY5Y cells (Chang et al., 2016 (link)). The expression of Aβ-GFP RNA increased 28-30 folds with the addition of 5 µg/mL of doxycycline (Sigma-Aldrich) (Huang et al., 2021 (link)).
Protocol detail
Alzheimer's Aβ Folding Reporter Assay
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A gene
Apigenin
Cell culture
Cells
Culture medium
Doxycycline
Flavones
Green fluorescent protein
Human
Pentahydroxyflavone
Quercetin
Rna expression
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Variable analysis
independent variables
- 7,8-DHF
- Quercetin (3,3',4',5,7-pentahydroxyflavone)
- Apigenin (4',5,7-trihydroxyflavone)
- Expression of Aβ-GFP RNA
- Cell culture medium
- SH-SY5Y cells (ATCC No. CRL-2266) with a single copy of the Aβ-green fluorescent protein (GFP) gene integrated
- 5 µg/mL of doxycycline (Sigma-Aldrich)
- Positive control: Addition of 5 µg/mL of doxycycline to increase the expression of Aβ-GFP RNA
- Negative control: Not explicitly mentioned
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