Fig. 3

Alkaline phosphatase staining, karyotype analysis, and embryoid body (EB) formation assay. (A). Feline iPSC colonies were stained for alkaline phosphatase or left unstained (control). (B). Karyotype analysis of feline iPSCs-1 and iPSCs − 2 at P20 showing a normal diploid chromosome number of 38. (C). Feline iPSCs formed embryoid bodies (EBs) in suspension culture in differentiation medium. (D). Conventional RT-PCR analysis of feline markers of all three embryonic germ layers, including alpha-fetoprotein (AFP), GATA binding protein 6 (GATA6), and C-X-C chemokine receptor type 4 (CXCR4) for endoderm; smooth muscle actin (SMA) and GATA2 for mesoderm; and ENOLASE and NESTIN for ectoderm in feline EBs. Feline GAPDH was included as loading control. Full-length gels are presented in Additional File 10: Fig. S10. (E). Conventional RT-PCR analysis of endogenously expressed feline pluripotency markers OCT4, SOX2, and NANOG, and the loading control GAPDH, in feline iPSCs and EBs