Fig. 2.

Translatome analysis reveals significant upregulation of neuronal mRNA in the HCs of GFI1 mutant mice. (A) P0 cochlear and vestibular tissues from mice expressing Rpl22^HA in HCs were collected separately for RiboTag immunoprecipitation. (B,C) In both cochlear (B, Gfi1^cre/+; B', Gfi1^cre/cre) and vestibular (C, Gfi1^cre/+; C', Gfi1^cre/cre) samples, immunoprecipitates (IPs) had higher levels of transcripts for the HC-expressed gene Myo6 compared with input (IN), but had lower levels of the transcripts for the mesenchymal-expressed gene Pou3f4. Immunoprecipitates were not enriched with Tubb3 (n=3). Dots represent individual replicates. Data are mean±s.d. (D) Tubb3 is upregulated in the mutant cochlear IPs compared with control IPs (fold change=7.87, P=0.0046), but not in mutant cochlear IN compared with control IN (n=3). (E) Number of genes upregulated and downregulated in the Gfi1^cre/cre cochlear and vestibular HCs. (F-H) Top 15 enriched gene ontology (GO) terms from genes downregulated (F) or upregulated (G) in Gfi1^cre/cre cochlear HCs, or upregulated in Gfi1^cre/cre vestibular HCs (H). (I-K) qPCR validation of dysregulated genes in vestibular (I) and cochlear (J,K) Gfi1^cre RiboTag immunoprecipitation samples (n=3). *P<0.05, **P<0.01, ***P<0.001, ns, not significant. Statistical significance assessed by a two-tailed Welch's t-test. Data are mean±s.d.