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. 2016 Jun 23;6:28532. doi: 10.1038/srep28532

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(a) Time course of the expression of different proteins in NT2 cells induced to differentiate with retinoic acid (RA). At each time point cells were lysed and the extracts were loaded in 10% acrylamide gel and analyzed by immunoblots. The quantification of the proteins in the immunoblots is shown in Supplementary Fig. S9. (b) The expression of Sox2 and VRK1 proteins in NT2 cells was detected by immunofluorescence before and after induction of differentiation with retinoic acid (RA). Sox2 was detected using a mouse mAb anti-Sox2 (E-4, 1:100). VRK1 was detected using a rabbit polyclonal anti-VRK1 (1:200) antibody. (c) Effect of RA-induced differentiation of NT2 cells on the gene expression levels Sox2, Oct4, VRK1 and macro H2A2. Total RNA was extracted at the indicated times and the RT-PCR was performed using specific primers indicated in the methods section.