*4.3. Cell Culture*

*PC12* (ATCC® CRL-1721™) rat cell line were used for experiments. Cells were cultured in DMEM, supplemented with 10% horse serum, 5% FBS, 1% L glutamine (200 mM), 1% of mixture penicillin (100 IU/mL), and streptomycin (100 μg/mL), at 37 ◦C in a CO2 incubator. Before the experiments, cells were differentiated by culturing in serum-free medium containing 50 ng/mL nerve growth factor (NGF) for 5 days. Protein concentration was estimated using the Bradford assay by spectrophotometrically reading at 595 nm [64].
