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

Figure 9. RNAi-mediated knockdown of p66Shc increases MnSOD levels and reduces superoxide production in early bovine embryos.

Figure 9

Bovine zygotes were either non-injected (controls), injected with negative control siRNA molecules, or injected with p66Shc-specific siRNAs (RNAi-E). Groups of embryos were either stained for MitoSOX™ detection of superoxide levels or immunostained for MnSOD protein content for quantification by relative fluorescent signal intensities. (A) Representative confocal images of 5–8 cell bovine embryos immunostained for MnSOD protein. (B) Embryos injected with p66Shc siRNA molecule RNAi-E exhibited significantly higher levels of MnSOD protein than control groups. (C) Representative confocal images of 2–4 cell bovine embryos were subjected to MitoSOX™ staining for superoxide quantification by relative fluorescent signal intensity. (D) Embryos injected with p66Shc RNAi molecules exhibited significantly lower levels of superoxide content than control groups. Different letters above the histogram bars represent significant differences (P<0.05) in mean fluorescent intensities. Scale bar, 50 µM.