Figure 8: For expression analysis, cultured OECs (NOECs (day 13); HOECs (day 30) were cultured in in T-25 flask in 1:1 Dulbecco’s modified Eagle’s medium (DMEM)/ Ham’s F12 containing penicillin, streptomycin, gentamicin, bovine insulin, Human epidermal growth factor (hEGF), Transforming Growth Factor-α (TGF-α), cholera toxin, hydrocortisone, Non-essential amino acids (NEAAs), 10% FCS and 10 mM HEPES in a humidified 5% CO2 incubator at 37°C. 30 μg of the whole cell lysate was separated on a 12.5% SDS-PAGE gel, probed with GFAP, p75NTR, caspase 3, 8, 9 and phospho-p53 antibodies following transfer to a nitrocellulose membrane. β-actin was used as a protein loading control. Data shown is representative of one of the three similar experiments each performed in triplicate.