3d molecular interactions visualized using biovia discovery studio visualizer v24.1.0.23298  (Accelrys)

Journal: EMBO Molecular Medicine

Article Title: AAV gene therapy rescues hearing and balance in a model of CLIC5 deafness

doi: 10.1038/s44321-025-00275-7

Figure Lengend Snippet: ( A ) 3D surface projection of IHCs and OHCs from the cochlear 32 kHz region of Clic5 +/− , Clic5 −/− and Clic5 −/− injected with scAAV. Clic5 at 12 weeks, stained with a CLIC5 (red) and actin (gray). ( B ) Representative confocal images of IHCs from the apical turn of Clic5 +/− and Clic5 −/− injected with scAAV. Clic5 at 12 weeks, stained with a CLIC5 (red), actin (gray), and FLAG tag (green). ( C ) Representative confocal images of IHCs and OHCs from the cochlear 8 kHz region of Clic5 +/− , Clic5 −/− , and Clic5 −/− injected with scAAV. Clic5 at 12 weeks, stained with actin. ( D ) Stereocilia length measurements of IHC at different regions of the cochlea, n = 3 for Clic5 +/− , Clic5 −/− , and Clic5 −/− injected with scAAV. Clic5 . ( E ) Quantification of OHC survival at different regions of the cochlea, n = 3 for Clic5 +/− , Clic5 −/− , and Clic5 −/− injected with scAAV. Clic5 . ( F ) Quantification of IHC survival at different regions of the cochlea, n = 3 for Clic5 +/− , Clic5 −/− , and Clic5 −/− injected with scAAV. Clic5 . ( G ) High-magnification scanning electron microscopy images of outer and inner hair bundles from the cochlea middle-turn of the Clic5 −/− and Clic5 −/− injected with scAAV. Clic5 at 12 weeks. ( H ) Representative images of IHCs and OHCs at 12 weeks at different regions of the cochlea of Clic5 −/ − injected with scAAV. Clic5 . ( I ) ABR thresholds at 4 weeks of Clic5 +/− ( n = 11), Clic5 −/− ( n = 10), and Clic5 −/− injected with scAAV. Clic5 ( n = 10). ( J ) ABR thresholds at 8 weeks of Clic5 +/− ( n = 11), Clic5 −/− ( n = 11), and Clic5 −/− injected with scAVV. Clic5 (n = 7). ( K ) ABR thresholds at 12 weeks of Clic5 +/− ( n = 10), Clic5 −/− ( n = 10), and Clic5 −/− injected with scAAV. Clic5 ( n = 7). ( L ) DPOAE thresholds at 4 weeks of Clic5 +/− ( n = 12), Clic5 −/ − ( n = 12), and Clic5 −/− injected with scAAV. Clic5 ( n = 10). ( M ) ABR thresholds at 3-time points of Clic5 −/− injected with scAAV. Clic5: 4 weeks ( n = 10), 8 weeks ( n = 7), and 12 weeks ( n = 7). ( N ) Freezing behavior duration in a fear conditioning assay at 12 weeks of Clic5 +/− ( n = 14), Clic5 − /− ( n = 14), and Clic5 −/− injected with scAAV. Clic5 ( n = 6). ( O ) Expression rates of scAAV. Clic5 across the cochlea as a function of viral titer, n = 3 for each group. ( P ) ABR thresholds at 4 weeks of Clic5 −/ − injected with scAAV .Clic5 at different viral doses: 1.52 × 10¹³ gc/ml ( n = 10), 1.52 × 10 12 gc/ml ( n = 3) and 1.52 × 10 11 gc/ml ( n = 3). Data information: Statistical tests were two-way ANOVA with Holm–Sidak correction for multiple comparisons for ( D – F , I – N , P ), and one-way ANOVA followed by Tukey correction for multiple comparisons for ( O ). Plots show mean ± SD. ns not significant, * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001 (Exact P values are provided in Appendix Table S ). Scale bars = 10 μm for ( A , C , H ), 2 μm for ( B ), 0.5 μm for OHC ( G ), and 2 μm for IHC ( G ). .

Article Snippet: Imaris 3D Interactive Microscopy Visualization software , Oxford Instruments . , version 10.1.

Techniques: Injection, Staining, FLAG-tag, Electron Microscopy, Expressing