|
Fig. 3 zfPGRN-A knockdown, rescue and over-expression. (A) Dose-dependent efficacy of morpholino (MO2) targeting the 5′ UTR of zfPGRN-A was assessed by Western blot analysis in comparison to its co-orthologue zfPGRN-B and actin. zfPGRN-A and B are represented by single bands of approximately 55 and 30 kDa, respectively. The Western blot analysis of MO knockdown efficacy is a representative of three independent injection sets. (B) Western blot analysis of protein extracts from embryos injected with 10 ng of zfPGRN-A MO1, 10 ng of zfPGRN-A MO2, zfPGRN-A-MO2 together with zfPGRN-A mRNA. Injection of 10 ng/embryo of the zfPGRN-A MO1 and MO2 effectively decreased zfPGRN-A translation, zfPGRN-A-MO together with mRNA resulted in normal zfPGRN-A translation. (C) Control experiments showing the results of the injection of 100pg hPGRN mRNA and 1ng gfp mRNA demonstrating translation into protein for both.