Western blotting
Total protein was extracted using RIPA lysis buffer (Applygen, Beijing,
China). A total of 40 μg of protein was loaded into a 10% sodium
dodecyl sulphate-polyacrylamide gel and electrophoretically transferred
to polyvinylidene fluoride membranes (Merck Millipore, Billerica, MA,
USA). After blocking with 5% milk for 1 h, the membranes were incubated
with specific antibodies at the indicated dilutions for 12–16 h at 4°C.
The membranes were scanned using an Odyssey Sa Imaging System (LICOR
Biosciences, Lincoln, NE, USA). The antibody information used in this
study was shown in the supplementary material, Table 2.