Birc3

After treatment, cells were washed twice with ice-cold phosphate-buffered saline (PBS) (Nissui Pharmaceutical, Tokyo, Japan) and harvested in RIPA buffer [25 mM Tris (pH 7.6), 150 mM NaCl, 1% NP-40, 1% sodium deoxycholate 0.1% SDS; Thermo Fisher Scientific, Waltham, MA, USA]. Protein concentrations were measured using the BCA protein assay kit (Thermo Fisher Scientific). Protein samples were separated on SDS-polyacrylamide gels and transferred to a polyvinylidene fluoride membrane. The membrane was probed with primary antibodies and subsequently probed with horseradish peroxidase-conjugating (HRP) secondary antibodies (1:10000; GE Healthcare, Little Chalfont, UK). Proteins were detected by enhanced chemiluminescence using ImmunoStar® Zeta (Wako Pure Chemical Industries). The chemiluminescence images were taken using the LAS-500 or LAS-4000 (GE Healthcare) device. Primary antibodies were purchased as follows: GAPDH (1:2000) from American Research Products (Waltham, MA, USA); Lamin A/C (1:1000), ARNT (1:1000), BIRC3 (1:1000), caspase-3 (1:1000) and cleaved caspase-3 (1:1000) from Cell Signaling Technology (Danvers, MA, USA); and β-actin (1:1000) from Sigma-Aldrich.