Figure 4
Live imaging for osteoclasts.
Evaluation of TRAP-GFP (a–c) and MMP9-DsRed (d–f) fluorescent signals. (a,b,d,e) Stacked ventral images in the pharyngeal region at day 4 (TRAP-GFP) and at day 6 (MMP9-DsRed) for the ground (a,d) and flight (b,e) groups. The images were captured under the same conditions between flight and ground samples shown in Supplementary Table S2 and constructed by Z-projection by use of ImageJ software. Yellow arrows point to bones with osteoclasts localized on them. up: upper pharyngeal bone, lp: lower pharyngeal bone. Scale bars = 50 μm. (c,f) The fluorescence intensity for TRAP (d) and for MMP9 (f) increased during the flight at days 4 and 6 of the observation period. Numbers of TRAP line fish: ground, n = 14 (day 4), 16 (day 6); flight, n = 9 (day 4), 7 (day 6). Numbers of MMP9 line fish: ground, n = 6 (day 4), 6 (day 6); flight, n = 4 (day 4), 3 (day 6). These numbers are shown in Supplementary Table S1. *P < 0.05, Student’s t-test. Error bars, s.e.m.
