Figure 5.
Dynamin–Gox interaction. (A) Temperature-shift protocol for inactivating dynamin in shits2 mutants. (B) PMI in control and shits2 mutants olf hair cells reconstructed by FIB-SEM. (C) PMI neck width measured from three independently reconstructed olf hair cells per genotype. Data were plotted as violin plots showing the distribution and median values. (shits2/y w: 118 points from three individual olfs; shits2/Y: 131 points from three individual olfs). Statistical significance was determined using the Mann–Whitney U test (two-sided), **: P < 0.001. (D) Number of PMIs quantified from FIB-SEM reconstructions of three olf hair cells per genotype. Counts were made from three independently reconstructed olf hair cells for each genotype (y w and shits2/Y). No formal statistical test was applied because the data represent simple counts. (E) TEM views of shits2/y w (control) and shits2/Y (mutant). (F) Quantification of envelope curvature. Point curvatures were calculated using the Kappa plugin in Fiji, and the median curvature of each envelope fragment was used as a representative value. Data were compared using the Mann–Whitney U test (two-sided). **: P < 0.01. (G) Maxillary palp of shits2/y w (control) and shits2/Y (mutant) stained for expression of HA–Gox and Ref(2)P. Asterisk indicates completely internalized olf hair cells. (H) Enlarged views of shits2/Y olf hair cells, showing normal (top and lower-left) and partially invaginated (lower right) phenotypes. (I) FIB-SEM view of plasma membrane and ER of shits2/Y olf #1. (J) Relationship of ER and plasma membrane in shits2/Y and y w (control). (K) ER–plasma membrane contacts in shits2/Y. (L) Distribution of ER–plasma membrane contact site (yellow dot) in shits2/Y olf #1. (M) Model of plasma membrane and ER interaction underlying nanopore formation. PMI formation is sustained by the stimulation by Gox and the clearance by dynamin. This dynamic interaction is coupled to ER-phagy, which supplies excessive lipids to the plasma membrane that buckles under the confinement by apical ECM. Envelope formation follows plasma membrane curvature. Bar: 100 nm (approx.), 1 µm (E, left), 200 nm (E, right), 10 µm (G), 1 µm (H, I), 200 nm (J, approx.), 100 nm (K, approx.), and 1 µm (L, approx.).

Dynamin–Gox interaction. (A) Temperature-shift protocol for inactivating dynamin in shits2 mutants. (B) PMI in control and shits2 mutants olf hair cells reconstructed by FIB-SEM. (C) PMI neck width measured from three independently reconstructed olf hair cells per genotype. Data were plotted as violin plots showing the distribution and median values. (shits2/y w: 118 points from three individual olfs; shits2/Y: 131 points from three individual olfs). Statistical significance was determined using the Mann–Whitney U test (two-sided), **: P < 0.001. (D) Number of PMIs quantified from FIB-SEM reconstructions of three olf hair cells per genotype. Counts were made from three independently reconstructed olf hair cells for each genotype (y w and shits2/Y). No formal statistical test was applied because the data represent simple counts. (E) TEM views of shits2/y w (control) and shits2/Y (mutant). (F) Quantification of envelope curvature. Point curvatures were calculated using the Kappa plugin in Fiji, and the median curvature of each envelope fragment was used as a representative value. Data were compared using the Mann–Whitney U test (two-sided). **: P < 0.01. (G) Maxillary palp of shits2/y w (control) and shits2/Y (mutant) stained for expression of HA–Gox and Ref(2)P. Asterisk indicates completely internalized olf hair cells. (H) Enlarged views of shits2/Y olf hair cells, showing normal (top and lower-left) and partially invaginated (lower right) phenotypes. (I) FIB-SEM view of plasma membrane and ER of shits2/Y olf #1. (J) Relationship of ER and plasma membrane in shits2/Y and y w (control). (K) ER–plasma membrane contacts in shits2/Y. (L) Distribution of ER–plasma membrane contact site (yellow dot) in shits2/Y olf #1. (M) Model of plasma membrane and ER interaction underlying nanopore formation. PMI formation is sustained by the stimulation by Gox and the clearance by dynamin. This dynamic interaction is coupled to ER-phagy, which supplies excessive lipids to the plasma membrane that buckles under the confinement by apical ECM. Envelope formation follows plasma membrane curvature. Bar: 100 nm (approx.), 1 µm (E, left), 200 nm (E, right), 10 µm (G), 1 µm (H, I), 200 nm (J, approx.), 100 nm (K, approx.), and 1 µm (L, approx.).

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