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Fig. 2 Ribeye A-domains inconsistently localize to synaptic ribbons at 3 dpf and 5 dpf.
Representative confocal maximum-intensity projection images of immunolabel or GFP in lateral line NM hair cells. Scale bars: 3 μm (main panels), 1 μm (D, inset). (A–A′) Ribeye a (A-domain)-myc alone (A) and merged with Ribeye b labeling synaptic ribbons (A′) in posterior lateral line NM1 hair cells of a 3 dpf larva. (B) Ribeye a (A-domain)-myc (cyan) and Vglut3 (magenta) immunolabel in a cross-section of an anterior lateral line NM at 5 dpf. (C–C′) Ribeye b (A-domain)-GFP alone (C) and merged with Ribeye a immunolabel (C′) in NM1 hair cells of a 3 dpf larva. White boxes indicate regions used in (D–D′′). (D–D′′) Insets of region in C′ showing basally-localized Ribeye b (A-domain)-GFP in relation to synaptic ribbons labeled with an antibody to Ribeye a. (D) Two adjacent hair cells: one with low Ribeye b (A-domain) expression next to another hair cell with moderate expression. A domain-GFP colocalizes with endogenous Ribeye a. (D′) A hair cell with moderate Ribeye b (A-domain) expression. (A domain)-GFP colocalizes with endogenous Ribeye a. (D′′) A hair cell with high Ribeye b (A-domain) expression. Read arrows indicate basally localized A-domain aggregates that do not colocalize with endogenous Ribeye. (E–E′) Ribeye a (A-domain)-myc (E), Ribeye b (A-domain)-GFP (E′), and merged with Ribeye b immunolabel (E′′) in NM1 hair cells of a 3 dpf larva. Note that the Ribeye b antibody labels both endogenous Ribeye b and exogenous Ribeye b (A-domain).