International Journal of Stem Cells : eISSN 2005-5447

Fig. 3.

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Fig. 3. Over-expression of Mbd3b facilitates reprogramming. (A) Immunoblot analysis for cells over-expressing Mbd3b compared to sham. Densitometry plot showed that cells over-expressing Mbd3b increased its expression by almost 50% compared to sham (p<0.05). (B) Quantification of iPSC colonies generated from fibroblasts transfected with RF+Mbd3b compared to cells treated with RF in BJ neonatal cell line and patient fibroblasts (RPD1 and PD1). A two fold increase is seen in the number of colonies in RF+Mbd3b compared to RF only (p<0.05). (C) Representative phase images of iPSC of established cell lines transfected with RF and RF+Mbd3b. Immunostaining with pluripotency markers (green) and counter staining with DAPI (blue) of cells treated with RF and RF+Mbd3b with TRA-1-81 and TRA-1-60. Scale bar is 200μm. (Error bars±S.E.M). (D) Q-PCR showed expression of pluripotency genes (Oct-4, Nanog and Sox-2 respectively) in cells treated with RF and RF+Mbd3b. All results were normalised to GAPDH (n=3 Error bars±S.E.M). (E) Representative images of embryoid bodies (EBs) generated from RF (left), and RF+Mbd3b (right). (F) Q-PCR showed up-regulation of lineage specific genes: Cdx2 (mesoderm), Pax6 (ectoderm) and MixI (endoderm) in conditions treated with RF EBs and RF+Mbd3b EBs (G) relative to respective iPSC line.
International Journal of Stem Cells 2018;11:235-41
© 2018 International Journal of Stem Cells