Journal: bioRxiv

Article Title: Single-Cell Profiling of the Developing Organ of Corti Identifies Etv4/5/1 as Key Regulators of Pillar Cell Identity

doi: 10.64898/2026.01.19.700450

Figure Lengend Snippet: a. List of top ten regulons (based on RSS) from SCENIC+ analysis of OC development. IPCs express five regulons (in green) that were not among the top ten regulons for any other cell type. b. Feature plots showing AUC values for Etv4 and Etv5 regulons. Both are increased in the IPC cluster (orange circles). c. Feature plots showing AUC values for Nkx6-1, Zfp410 and Zfp420. None show obvious elevation in the IPC cluster. d. Etv gene network developed by combining the list of target genes in the Etv regulons with the DE gene list for IPCs. e. Violin plots for the 14 genes listed in d. Four are IPC-specific (orange). f. CellOracle analysis. Upper panel: GRN signal propagation arrows indicated developmental flow along the medial and lateral trajectories. Perturbations: Upper panels, arrows indicate predicted changes in developmental trajectories for three genes known to be involved in cochlear development, Atoh1, Pou4f3, Hes5 . Lower panels, subtraction of perturbed vector flow from GRN signal propagation indicates regions of developmental enhancement (green) or inhibition (magenta). g. CellOracle analysis for perturbation of Etv4 . Results indicate inhibition of IPC and Lat.PsC development and possible enhancement of OPC/DC and OHC development.

Article Snippet: Etv1 flox , Etv4 - and Etv5 flox mice were obtained from MMRRC and maintained in a mixed genetic background – .

Techniques: Plasmid Preparation, Inhibition

Journal: bioRxiv

Article Title: Single-Cell Profiling of the Developing Organ of Corti Identifies Etv4/5/1 as Key Regulators of Pillar Cell Identity

doi: 10.64898/2026.01.19.700450

Figure Lengend Snippet: a. Surface views of the indicated regions of the OC in control, Etv5/1 double KO and Etv Triple KOs at E18. In control a single row of IHCs is separated from three rows of OHCs by an ordered row of IPCs (NGFR in magenta). Etv double KO cochleae appear normal although some subtle patterning defects may be present in the apical OHC region. In Etv Triple KO cochleae, IPC development appears disrupted, NGFR expression is absent in most IPCs, and cellular development and patterning appears significantly disrupted in the apical region. In addition, small ectopic hair cells may be present in the basal turn in Etv triple Kos (arrows). b. Expression of a second IPC marker, NPY (green), is downregulated in both Etv double and Triple KOs. c. Quantification of HC density at different positions along the OC in control and Etv Triple KOs. Boxes indicate 1 st and 3 rd quartiles, interior line in each box indicates the mean, blue dots indicate individual data points. A significant decrease in OHC density is present in the middle and apical regions of the cochlea in Etv Triple KOs. d. Characterization of OHC patterning. The position of the six nearest OHCs were determined for OHCs located in the second row. Those positions were then plotted into 16 radial bins. The radar plot compares the frequency of OHCs in each radial bin in the basal region of the OC from control and Etv Triple KOs. Overall, Etv Triple KOs showed greater variability in the patterning of OHCs. Asterisks indicate sectors with significant changes between control (green) and Etv Triple KOs (magenta). * p<0.05; **, p<0.01. Scale bar in a (same in b) 20 μm. Abbr. Phal.: Phalloidin.

Article Snippet: Etv1 flox , Etv4 - and Etv5 flox mice were obtained from MMRRC and maintained in a mixed genetic background – .

Techniques: Control, Expressing, Marker