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Fig. 4
Microinjection of irx7SMO (10 ng) and irx7MO2 (3 ng) drastically reduces Irx7 protein level.
The heads of 35 embryos injected with control MO and irx7SMO, and 30 embryos injected with irx7MO2-6 bms and irx7MO2 were dissected at 72 hpf. Proteins were then extracted from these samples and Irx7 expression detected by Western blot using anti-Irx7-234 (A). Γ-tubulin was used as a loading control. The specificity of the Irx7 antibodies was first confirmed by Western blot using various recombinant Irx7 proteins of different lengths (Table S1 and File S3) expressed from bacterial culture (data not shown). Using the information extracted from the Western blot, the protein level of Irx7 was found to be reduced by 69.01% and 87.58% respectively in the dissected heads of irx7SMO and irx7MO2 morphants when compared to the corresponding controls (B).