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. 2015 May 27;5:10283. doi: 10.1038/srep10283

Figure 2.

Figure 2

A: Embryos were injected into both cells at the two cell stage with RNAs encoding GFP (150 pgs per embryo) either alone or together with Cetn2MO1, Cetn2MO2, or Cetn3MO (10 ngs/side, 20 ngs total per embryo); at stage 11 or 25 the embryos were analyzed by SDS-PAGE and immunoblot using the anti-Human Cetn-1 antibody (which reacts preferentially with Cetn2 compared to Cetn3. There was a clear decrease in Cetn2 protein levels, persisting through stage 25. To confirm the specificities of the Cetn MOs both blastomeres of two cell embryos were injected with RNAs encoding GFP (200 pg/side) and RNAs encoding Cetn2a-GFP (B), Cetn3l-GFP (C), or Cetn4-GFP (D) RNAs with (“+”) or without Cetn MO (10 ng/side). These Cetn RNAs contain the target sequence of the corresponding morpholino. In addition, uninjected (“UN”) and embryos injected with GFP RNA alone were examined as controls for antibody specificity. Injected embryos were harvested at stage 11. Immunoblot analyses were carried out using an anti-rabbit GFP antibody. An apparent breakdown product of the Cetn2-GFP construct is indicated by the arrow in the Cetn2 MO panel.