Breeding schedule and outcome of the generation of En1KO;Pitx3GFP/GFP animals.

(A) En1Het;Pitx3GFP/GFP animals were inter-crossed to generate a litter that includes three genotypes: En1WT;Pitx3GFP/GFP, En1Het;Pitx3GFP/GFP, and En1KO;Pitx3GFP/GFP. (B) The presence of double En1KO;Pitx3GFP/GFP animals occurred below (Mendelian) predicted chance level (12.7% versus 25%). (C) DAPI staining at E14.5 reveals that in En1WT;Pitx3GFP/GFP and En1Het;Pitx3GFP/GFP animals the cerebellar anlage is correctly developed (Cb), whilst it is absent in the double En1KO;Pitx3GFP/GFP (*). (Rf: retroflexus, Cb: cerebellum, Cp: choroid plexus).