This record contains data (Table S1 to S3, Figure S1 to S12, File S1) related to article “In-depth genetic and molecular characterization of Diaphanous Related Formin 2 (DIAPH2) as a novel candidate gene for hearing loss” . Specifically, the record contains the following supplementary information to the article: Table S1. Coding sequence coverage of known autosomal recessive and X-linked NSHL-causing genes in exome data. Table S2. Prioritized variants shared between affected siblings III1 and III3. Table S3. Primers used for genetic screening of candidate genes/variants. Figure S1. Analysis of auditory brainstem evoked potentials in proband III3. Figure S2. Expression of mouse Diaph2 mRNA in P4 organ of Corti by RT-PCR. Figure S3. Diaph2 expression in E14.5 and E16.5 wild-type mouse cochlea. Figure S4. Evaluation of Diaph2 expression in whole-mount mouse cochleas. Figure S5. Diaph2 expression in P7 and P14 wild-type mouse cochlea. Figure S6. In-silico analysis of the impact of c.868A>G variant on DIAPH2 pre-mRNA splicing. Figure S7. In-vitro analysis of the impact of c.868A>G variant on DIAPH2 pre-mRNA splicing. Figure S8. In-vitro characterization of the effect of the c.868A>G variation on splicing using a DIAPH2minigene spanning exons 6 to 9. Figure S9. Analysis of DIAPH2 exon 8 splicing in blood from NSHL3 family subjects. Figure S10. In-vivo analysis of Diaph2 exon 8 splicing in the mouse cochlea. Figure S11. Pathogenicity prediction of the p.I290V missense variant with 8 commonly used software. Figure S12. DIAPH2 immunolocalization studies in basal conditions. File S1. Supplementary Methods