Figure 5

Microinjection of miR-196a mimic represses endogenous NOBOX expression in bovine early embryos. (A) Effect of miR-196a mimic microinjection on abundance of NOBOX protein in 8-cell stage embryos as determined by immunocytochemical analysis using confocal spinning-disk microscopy (n = 4 pools of 5-10 embryos per treatment). Uninjected embryos and embryos injected with a nonspecific miRNA (Neg miRNA) were used as controls. Nuclear DNA was stained with DAPI. (B) Effect of miR-196a mimic microinjection on abundance of NOBOX mRNA in eight-cell embryos as determined by real-time PCR. Data were normalized relative to abundance of endogenous control ribosomal protein S18 (RSP18) and are shown as mean ± SEM (n = 4 pools of 10 embryos per treatment). Different letters indicate statistical difference (P < 0.05).