Immunolocalization and PCR of FSHR and OCT-4 in the granulosa cells, oocytes and embryo. ( A ) Confocal microscopy shows the presence of ( a ) FSH receptors and ( b ) OCT-4 in the cytoplasm of the granulosa cells ( c-f ) OCT-4 localization in MI and MII oocytes and early embryo. Note the MI oocyte shows both nuclear and cytoplasmic OCT-4 whereas in the MII oocyte OCT-4 was observed in the ooplasm in the 4–8 cells embryo and blastocyst OCT-4 was observed in the blastomeres surrounding trophoectodermal cells as well as in the inner cell mass. ( B ) qRT-PCR analysis of FSH receptor transcripts R1 and R3, Oct-4A and total Oct-4 in the granulosa cells surrounding MI and MII oocytes. R3 transcript mRNA is significantly highly expressed compared to the canonical R1 transcript in the granulosa cells. Note a reduction in all the transcripts in the granulosa cells collected from MII oocytes. Magnification is 40X and granulosa cells are image at 4X optical zoom. The relative expression of transcripts is a representation of pooled samples analyzed once.