XB-IMG-82245
Xenbase Image ID: 82245
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FIGURE 6. MO-mediated knockdown of endogenous rdhe2. A, Western blot analysis of protein extracts (30 μg/lane) from neurula embryos injected with 4 ng of rdhe2 mRNA alone or mixed with 50 ng of rdhe2 MO. MO essentially prevents an increase in the newly synthesized rdhe2 protein levels over those preexisting in wild-type embryos (bottom). Top, staining for β-actin as a loading control. B, Western blot analysis of protein extracts prepared from an equal number of rdhe2 and control morphants (st. 32) using rdhe2 (bottom) or actin (top) antibodies shows a decrease in endogenous rdhe2 levels. C, array of phenotypes displayed by rdhe2 and rdh10 morphants compared with control morphants. Phenotypes were divided into severe, strong, moderate, and mild groups with the representative embryos shown for each group. An arrow shows an incompletely closed neural tube typical for severely affected rdhe2 morphants. Dramatic phenotypic changes in morphants compared with the moderate decrease in protein levels in B may reflect a strong localized effect of MO at sites where newly synthesized rdhe2 protein is critical for embryogenesis. D, proportion of different phenotypic groups in rdhe2 morphants compared with control morphants and wild-type embryos. E, co-injection of rdhe2 rescue mRNA (4 ng) and rdhe2 MO (30 ng) decreasing the frequency and severity of defects displayed by morphants. F, proportion of different phenotypic groups in embryos injected with 25 ng of rdhe2 MO or rdh10 MO, or a mixture of both (50 ng total). Control MO (25 ng) was added to individual rdhe2- and rdh10-MO injections to equalize the total oligonucleotide amount. Image published in: Belyaeva OV et al. (2012) Copyright © 2012. Image reproduced with permission of the publisher and the copyright holder. This is an Open Access article distributed under the terms of the Creative Commons Attribution License.
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