Luteolin and Oxaliplatin Cytotoxicity Assay

A total of 3×10⁶ MFC cells were inoculated into 100-mm culture dishes. After overnight culture, the cells were incubated with luteolin (20 µM) and/or oxaliplatin (5 µM) (total volume 7 ml/well) for 24 h. Then, the cells were rinsed with PBS and lysed on ice for 30 min with RIPA buffer (cat. no. R0010; Beijing Solarbio Science & Technology Co., Ltd.) containing 0.1 M PMSF (cat. no. P0100; Beijing Solarbio Science & Technology Co., Ltd.) and protease phosphatase inhibitor (cat. no. P1261; Beijing Solarbio Science & Technology Co., Ltd.). The cell lysate was centrifuged at 12,000 × g (Thermo Scientific™ Sorvall™ Legend™ Micro 21R Microcentrifuge; Thermo Fisher Scientific, Inc.) for 15 min at 4°C, and the supernatant was collected for the subsequent tests. The protein quantification was performed using the BCA protein assay kit (cat. no. PC0020; Beijing Solarbio Science & Technology Co., Ltd.) (30 (link)). The data were obtained at the absorbance of 562 nm by a microplate reader (Infinite 200 PRO, Tecan, Tecan Austria GmbH). The amount of protein was calculated in accordance with the prescribed computational formula of the kit's protocol.