Murine Fam65b forms ring-like structures at the base of stereocilia critical for mechanosensory hair cell function
- The Scripps Research Institute, United States
Figures
Figure 1
Analysis of hearing function in Fam65b-deficient mice.
(A) Diagram of cochlear hair cells showing the mechanically sensitive hair bundle. Stereocilia are arranged into three rows of decreasing heights. Stereocilia form a taper at their base near the …
Figure 2
Expression of Fam65b in hair cells.
(A–C) Cochlear whole mounts from P4 Fam65bLacZ/+ mice were stained for LacZ. (A) Whole mount staining reveals expression of Fam65b along the length of the cochlear duct. (B) Higher magnification …
Figure 3
Hair cell morphology in Fam65b-deficient mice.
(A, C) Analysis of the cochlea in P2 (A) and P5 (C) control and mutant animals by whole mount staining with phalloidin (green). Note the morphological changes in hair bundles from mutant mice with …
Figure 4 with 1 supplement
Analysis of hair bundle morphology by SEM.
Whole mounts from the middle part of the cochlea from Fam65bLacZ/+controls and Fam65bLacZ/LacZmutants at P5(A) where analyzed by SEM. (A) Low magnification view showing disorganization of hair …
Figure 4—figure supplement 1
Lack of Fam65b interactions with taperin.
(A) Co–immunoprecipitation experiments. CL4 cells were transfected with the constructs indicated on top of each panel. Immunoprecipitations were carried out with HA antibody followed by western …
Figure 5
Taperin localization and shRNA perturbation of Fam65b.
(A, B) Taperin (green, A) and espin (green, B) staining in control and mutant IHCs analyzed by fluorescent deconvolution microscopy. Stereocilia were visualized by staining with phalloidin–rhodamine …
Figure 6
Analysis of mechanotransduction currents in Fam65b-deficient hair cells.
(A) Examples of transduction currents in OHCs from control and mutant mice at P5 in response to a set of 10 msec hair bundle deflections ranging from –400 nm to 1000 nm (100 nm steps). (B) Current …
Figure 7 with 1 supplement
Distinct localization pattern of Fam65b and taperin resolved by STORM.
(A–F) STORM images of Fam65b (A–C) and taperin (D–F) localization in hair cells. A and D show magnification/resolution similar to fluorescent deconvolution microscopy or traditional confocal …
Figure 7—figure supplement 1
Minimal hair cell loss in Fam65b mutants at P16 and P28.
(A,B) SEM images of the middle part of the cochlea from P16 (A) and P28 (B) Fam65Lacz/+control (left) and Fam65Lacz/Laczmutants (right). Although hair bundles were disorganized at P16 and P28 in Fam6…
Figure 8
Fam65b forms oligomeric structures.
(A) Diagram of the constructs used for biochemical experiments. Evolutionarily conserved domains are indicated in black. (B–G) HEK293 cells were transfected with the constructs indicated on top of …
Figure 9
Effects of RhoC on Fam65b oligomerization.
(A) HEK293 cells were transfected with the constructs indicated on top of each panel. Immunoprecipitations were carried out with HA antibody, followed by western blotting to detect co–expressed …
