RiboTag analysis of Gfi1Cre/Cre and Gfi1Cre/+ cochlear and vestibular hair cells

In order to identify the transcriptional cascade downstream of the GFI1 repressor in inner ear hair cells, we performed a RiboTag-based expression analysis of Gfi1cre/cre mutant hair cells compared to their heterozygous controls. Through this analysis, we identified a significant downregulation of genes involved in normal hair cell development and function within the mutant hair cells, suggesting that in the absence of GFI1, the hair cells undergo a maturation arrest. Furthermore, the Gfi1 deficient hair cells displayed a significant upregulation of genes involved in neurodifferentiation and neuronal function. Comparison of the P0-P1 hair cell translatome of Gfi1 mutants and heterozygous controls by HA-tagged ribosome immunoprecipitation and RNA-seq. Cochlear and vestibular immunoprecipitated samples were compared in independent biological triplicates.