Mutation of Growth Arrest Specific 8 Reveals a Role in Motile Cilia Function and Human Disease
Fig 1
Generation of mutant Gas8GT mice and phenotype description.
(A) Schematic of the wildtype Gas8 allele (Gas8WT) and the Gas8 genetrap allele (Gas8GT). The relative position of the β-geo cassette is indicated by the blue box. Arrows indicate the primers used for RT-PCR analysis. (B) RT-PCR expression analysis of Gas8 transcript in Gas8WT, Gas8GT/WT, Gas8GT whole embryos shows the presence of the 5’ end of Gas8 transcript (left panels) and absence of the 3’ end (right panels). Actin served as a positive template control in all samples. Reactions with reverse transcriptase are indicated (+) and negative RT controls (-). (C) Western blot for Gas8 protein on Gas8WT, Gas8GT/WT, and Gas8GT trachea. Wildtype Gas8 is located at 57 kDa (Gas8WT) while the genetrap allele is at approximately 230kDa (Gas8GT). * denotes a spurious band recognized by polyclonal antibody. GAPDH was used as a loading control. (D) Nissl stained coronal section of P21 Gas8WT and Gas8GT brain. (E) Gas8GT mice display situs inversus as noted by the reversed direction of the heart apex (white lines indicate heart axis) and (F) the stomach location in P2 pups (arrow).