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Western blot analysis

XC Xuzi Cai
KW Kang-Nan Wang
WM Wen Ma
YY Yuanyuan Yang
GC Gui Chen
HF Huijiao Fu
CC Chunhui Cui
ZY Zhiqiang Yu
XW Xuefeng Wang
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Skov3 cells were seeded in 6-well plates and cultured for 12 h. After incubated with Ir-1 (2.0 μM) and Ir-NPs (2.0 μM) for 12 h, cells were treated with or without white light (400–700 nm) irradiation at 50 mW/cm2 for 5 min. After 12 h of incubation, the cells were collected to extract protein with radioimmunoprecipitation assay lysis buffer. Equal amounts of these proteins, as determined by BCA Protein Assay Kit, were added to SDS-PAGE gels and separated by gel electrophoresis, respectively. After transferring protein from the gel to the polyvinylidene difluoride (PVDF) membrane, the membrane was blocked with 5% BSA, then incubated with primary antibodies (Bcl-2, Bax, Cytochrome c and Caspase-3) and Anti-β-actin rabbit monoclonal antibody. Subsequently, the membrane was incubated with goat anti-rabbit IgG antibody. The blots were exposed by an image analysis system (Bio-rad, USA).

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