Equal amounts of protein were loaded into the SDS-PAGE gels and then transferred to polyvinylidene fluoride (PVDF) mem- branes (Millipore, Boston, MA, USA). The membranes were blocked with 3% BSA and incubated with different primary antibodies at 4 °C overnight. The membranes were washed...
Then, 100 μL SDS-HCl was added to each well, and the plate was incubated overnight at 37 °C. The optical density (OD) was analyzed at 570 nm on a Microplate Reader (Thermo Skanit Software 3.2, Germany)16. Preparation of the subcellular fraction of the ER and mitochondria PC12 cells...