Antibodies and Western blot

Antibodies against AASS (1:1,000; HPA020728) and α-tubulin (1:5,000; T6199) were from Sigma-Aldrich. Western blot analysis was performed using liver homogenates. To prepare liver homogenates, 100 mg liver tissue was collected, snap-frozen in liquid nitrogen, homogenized, and ultrasonicated in 400 µl tissue lysis buffer (50 mM Tris-HCl, pH 7.5, 100 mM NaCl, 2 mM EDTA, and 1% NP-40) with protease inhibitor (05892970001; Roche) and phosphatase inhibitor (04906837001; Roche). Liver homogenates were collected from the supernatant after centrifugation at 12,000 g at 4°C for 10 min.