Figure 1—figure supplement 2.

Figure 1—figure supplement 2. Expression of Semaphorin7A in the lateral line neuromast.

(A–C) Surface micrographs show hair cells marked with actin-GFP (green dashed lines) and the localization of Sema7A protein (orange) in the subapical (arrowheads) and basal regions (arrows) of the corresponding hair cells from neuromasts of developing control larvae. The inset in panel A (red arrowheads) depicts the apices of the immature hair cell pair. (D) Immunolabeling with only the secondary antibody fails to detect any Sema7A signal. (E) A surface micrograph depicts hair cells (green, left) whose subapical region (red dashed line) harbors the Golgi network−labeled by the Golgi resident protein GM130 (cyan, middle)−where the Sema7A protein is enriched (orange, right). (F) A micrograph at a nuclear plane of a 4 dpf neuromast depicts membrane-tethered Scarlet (green) in all cells of the neuromast and the Sema7A protein (orange, arrowheads) in centrally clustered hair cells. (G) The Sema7A protein is located at the basal membrane (red arrowhead). (H) The Sema7A protein is also observed adjacent to the basal membrane, potentially in a vesicle (cyan arrowhead). (I) A plot quantifies the distribution of average Sema7A intensity across the hair cell’s basal membrane. The results stem from 100 hair-cell membranes of 10 neuromasts from 4 dpf larvae. (J) A schematic diagram of a single hair cell depicts the three distinct regions along the apicobasal axis of the cell. Sema7A intensity was measured along this apicobasal axis (red dashed line). Scale bars, 5 µm; au, arbitrary unit; means ± SEMs.