Fig 2: Immunofluorescence identification of the generated iPS. The newly derived mRNA iPS colonies were stained for the specialized markers of pluripotency: Oct4, Sox2, Nanog, SSEA-1, Klf4 and c-Myc. Nuclei were counter-stained with DAPI; (magnification 200×).

Fig 3: Plasmid construction and mRNA synthesis. (a) Successful amplification of the four factors from selected organs using specific primers for each. Oct4 (O) and Sox2 (S) were amplified from testes while c-Myc (C) was amplified from small intestine. Klf4 (K) was correctly amplified from colon. The DNA marker (M) used was a 2000 bp marker; (b) The eukaryotic expression vector (pCDNA 3) was digested using the same enzymes used for cutting the genes. Lane (1) shows the result of double restriction of the plasmid using EcoRI and XhoI, while Lane (2) revealed the single cut using XhoI. Lane (3), shows the uncut circular plasmid. (M): DNA marker (5000 bp); (c) Confirmation of the successful cloning through digestion of the newly formed recombinant plasmids (pCDNA–Oct4, pCDNA–Sox2, pCDNA–cMyc and pCDNA–Klf4) using EcoRI and XhoI enzymes resulted in two bands that were similar to the expected size of each gene as showed in Lane (2), while Lane (1) shows the single cut of each recombinant plasmid (M): DNA marker (5000 bp).