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Fig. 5 Methionine proximity-activated imaging reporter (Met-PAIR) staining on calmodulin (CaM) in zebrafish with external stimulation. Live zebrafish (3 day post-fertilization) were treated with Ox4 (100 μM) at rt for 20 min, further functionalized with biotin-azide via copper-catalyzed azide alkyne cycloaddition (CuAAC), and stained by mouse anti-CaM antibody and rabbit anti-biotin antibody as primary antibody and proximity-ligation assay (PLA) reagents. (a) Schematic illustration of the head of zebrafish for stimulation of posterior lateralis ganglion (PLLg) response through physical vibration. A: Anterior. D: Dorsal. P: Posterior. V: Ventral. (b) Representative confocal images of zebrafish stained by Met-PAIR with increases (vibration) or decreases (anesthesia with 920 μM tricaine) in local calcium. The images are representative of six biological replicates for each condition. PLA signal is shown in magenta. PLLg is noted with a red arrow. DAPI counterstaining and merged images are shown in Figure S17. Scale bar: 100 μm.