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MTT cell viability assay

SY Seung-Hoon Yang
JS Jisu Shin
NS Naewoo Neo Shin
JH Ji-Hyun Hwang
SH Sung-Chul Hong
KP Keunwan Park
JL Jae Wook Lee
SL Sejin Lee
SB Seungyeop Baek
KK Kyeonghwan Kim
IC Illhwan Cho
YK YoungSoo Kim
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To examine the effect of Nec-1 or Nec-1i on Aβ-induced brain cell death, HT22 and BV2 cells were assessed using MTT cytotoxicity assay as previously reported13. Cultured cells were plated into a 96-well plate (5 × 103 cells/well). Aβ42 solutions dissolved in DMSO (10 mM) were diluted 10 times using cell starvation medium (0.5% fetal bovine serum in DMEM) and then incubated for 24 hours at 37 °C for aggregation. After treatment of pre-aggregated Aβ42 (10 μM), Nec-1 (10, 25, 50, 100 μM), and demethylated Nec-1 (Nec-1i, 50 μM), MTT reagent (15 μM) was added to each well and was re-incubated for 4 more hours. Solubilization solution was then added and the plate was kept in room temperature overnight. The insoluble formazan was measured using SynergyTM HT Multi-Detection microplate reader (Bio-Tek).

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