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. 2014 Jan 24;9(1):e86978. doi: 10.1371/journal.pone.0086978

Figure 1. Real-time PCR quantification of p66Shc mRNA levels following RNAi-mediated knockdown of p66Shc in early bovine embryos.

Figure 1

Total RNA was extracted from pooled groups of 2–4 cell embryos that, at the zygote stage, were either non-injected, injected with negative control short interfering (si)RNAs, or injected with the p66Shc-specific siRNA molecules (RNAi-A or RNAi-E). (A) Real-time PCR quantification of p66Shc mRNA levels following a titration series of short interfering (si)RNAi-E molecules. The decrease in p66Shc mRNA levels became significant by the 0.05 µM dose level. (B) Histone H2A mRNA was quantified in parallel as an internal control of PCR efficiency. (C) Embryos injected with negative siRNA exhibited significantly higher levels of p66Shc mRNA than non-injected controls. Embryos injected with either RNAi-A or RNAi-E molecules exhibited significantly decreased abundance in p66Shc mRNA. The reduction induced by the RNAi-E molecule was significantly greater than that of the RNAi-A molecule. (D) Histone H2A mRNA was quantified in parallel as a control of PCR efficiency. Different letters above the histogram bars represent significant differences (P<0.05) in transcript abundance. No significant differences in H2A mRNA were noted between groups.