Two—three wells (each 3.8 cm2) of hNES cells, hiPSCs or neurons were collected at different time points (day 5, 8, 18 and 49) and RNA isolated using trizol extraction and iso-propanol precipitation method. Equal amounts of RNA per sample were used for cDNA synthesis (1 μg). The Q-PCR experiments were performed on Light cycler 480 (Roche) equipment. The expression of each marker during differentiation days 8, 18 and 49 is normalized to the expression of the same marker at day 5. Primers used for RT- and Q-PCR are listed in Table B in S1 File.
