Mouse Tissue Immunoblot Analysis

Mouse tissues were lysed with using RIPA buffer (Elpis-Biotech, Daejeon, Korea) and subjected to immunoblot analysis as described previously [46 (link)]. Proteins from liver whole and cytosolic fraction lysates were separated by 10% SDS- PAGE and transferred to nitrocellulose membranes. The membranes were probed with anti-β-actin (AbFrontier; diluted 1:5000) [26 (link)], anti-CYP2E1 (Proteintech, 19937-1-AP, 1:1000), anti-Cleaved caspase3 (Cell signaling, #9661; diluted 1:1000), anti-Cytochrome C (Cell signaling, #11940; diluted 1:1000), anti-smac (Abcam, ab32023; diluted 1:1000), and anti-α-tublin (AbFrontier; diluted 1:5000). The membranes were probed with specified antibody. Immunoreactive proteins were visualized using an Amersham ECL kit (GE Healthcare, Piscataway, NJ, USA) or using iBright CL1000 imaging system (Invitrogen) according to the manufacturer's instructions.