Fig. 4

Functional consequences of mutations in the MYH3 gene. a Result of the RT-PCR from CP and WT oculomotor muscles with primers MYH3ex21.1 and MYH3ex27.2 spanning exons 21 to 27 demonstrates that MYH3 mRNA expression is reduced in affected animals, but no alternative transcript resulting from the deletion could be identified. M marker, bp base pairs, WT wild-type, CP cleft palate. b Relative expression of MYH3 from CP and WT oculomotor muscles. The RT-qPCR of MYH3 exon 23 confirms the mRNA decay. The mRNA levels were normalized with the RPL13A and SDHA genes. WT wild-type, CP cleft palate. c Proteins were extracted from CP and WT oculomotor muscles. Samples were analysed by immunoblotting with an antibody against the MYH3 protein. At the protein level, the mutation led to a functional knockout as no MYH3 protein was identified by western blot in CP cases. M marker, kDa kilo Dalton, WT wild-type, CP cleft palate