Oxidative Medicine and Cellular Longevity / 2020 / Article / Fig 7

Research Article

New Synthetic 3-Benzoyl-5-Hydroxy-2H-Chromen-2-One (LM-031) Inhibits Polyglutamine Aggregation and Promotes Neurite Outgrowth through Enhancement of CREB, NRF2, and Reduction of AMPKα in SCA17 Cell Models

Figure 7

Trx-His-TBP/Q20−61 proteins and TBP aggregation monitored by thioflavin T fluorescence and filter trap assays. (a) SDS-PAGE gel of Trx-His-TBP/Q20−61 proteins from IPTG-induced bacterial cells (left) and Western blot analysis of purified TBP proteins using anti-TBP antibody (right). (b) Thioflavin T binding assay for Trx-His-TBP aggregation. TBP protein (5 μg in 200 μL PBS) was incubated with congo red (1−10 μM), LM-031, or licochalcone A (1−100 μM) at 37°C for 24 h, and aggregation was monitored by measuring thioflavin T fluorescence intensity (). values: comparisons between TBP/Q61 versus TBP/Q20 (###) or with and without compound addition (, , ). (c) Filter trap assay of TBP aggregation without or with congo red (10 μM), LM-031, or licochalcone A (100 μM) addition using TBP antibody (). To normalize, the relative trapped TBP/Q61 without compound addition is set as 100%. values: comparisons between TBP/Q61 versus TBP/Q20 (###) or with and without compound addition ().
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