Anti bcl2l10

Western blot was carried out as described in our previous study.28 (link) Briefly, total protein of HCC cells and tissues together with healthy liver tissues were lysed with RIPA lysate (containing protease inhibitors and phosphatase inhibitors, 1%) in order to extract the total protein. About 40 µg of total protein in each sample was added to a sample well of a 10% sodium dodecyl sulfate-polyacrylamide gel (SDS-PAGE) and electrophoresed. Separated proteins were then blotted onto a polyvinylidene fluoride membrane (EMD Millipore, Billerica, MA, USA). Membranes containing the imprinted protein were blocked with 0.5% bovine serum albumin for 2 hours, then the membrane was incubated with anti-Bcl2L10 (1:1,500; Cell Signaling Technology [CST], Danvers, MA, USA) or anti-β-actin (1:3,000; CST) antibodies overnight at 4°C. The next day, the membranes were rinsed thrice and further left to incubate with diluted secondary horseradish peroxidase-marked antibodies (1:3,000; CST) for 1 hour at room temperature. An enhanced chemiluminescence detection kit (EMD Millipore) was used to detect immunoreactive protein bands. Images were captured by ImageJ (National Institute of Mental Health, Bethesda, MR, USA).