Fig. 3: Loss of Jag1 reduces the expression of progenitor and SC-specific genes and attenuates PI3K-Akt-mTOR signaling.

a Experimental scheme. Cochlear epithelial cells from stage P2 mice were used to establish control and Jag1 cKO organoid cultures, which were analyzed on day 7 of expansion. b Volcano plot of RNA-seq data. Plotted is the beta-value (x-axis) versus −log10 q-value (y-axis). Transcripts that are significantly upregulated in response to Jag1 cKO are marked in red dots, and transcripts that are significantly downregulated are marked in blue dots. c Biological processes and pathways associated with Jag1 cKO downregulated genes ranked by adjusted p-value (q-value). d RT-qPCR was used to analyze relative mRNA expression of Jag1, Hey1, Id1, Eif4ebp1, Minar2, Nupr1, Dkk3, Scx in control and Jag1 cKO organoids (n = 4 animals per group, two independent experiments). e Immunoblots of p-Akt, Akt, p-S6, p-4EBP1, 4EBP1, JAG1 and β-actin proteins in control and Jag1 cKO organoids. f Normalized p-Akt, p-S6, and p-4EBP1 protein levels in (e) (n = 3 animals per group, from one representative experiment, two independent experiments). Graphed are individual data points and mean ± standard deviation. A two-tailed, unpaired Student’s t test was used to calculate exact p-values. Source data are provided as a Source Data file. N.s. not significant. Created in BioRender. Doetzlhofer, A. (2025) https://BioRender.com/vo09ioe.