waves. (a) Damage-associated propagating waves. The damaged site is indicated by a white arrowhead and a broken red circle. The pink arrowheads indicate the wing veins. The scale bar represents 300 μm. (b) The circular propagation of wave fronts from the damaged site shown in a from 4’00” to 4’40” (left panel) and from 9’20” to 11’20” (right panel). Three ROIs were set for analysis of intensity changes. See also Additional file 6: Movie S6. Damage-associated waves were induced successfully in 3 out of 9 treated individuals. (c) Fluorescence intensity changes over time in the 3 ROIs set in b. The identical waves are indicated by dashed lines. (d) Damaged wings with large elevations in Ca2+ signals around the damaged sites were observed in 6 out of 9 treated individuals. No generation of clear propagating Ca2+ waves was observed, but there were many small flickering waves around the damaged sites. ROIs were set for analysis of intensity changes. The scale bar represents 300 μm. (e) Fluorescence intensity changes over time in the 3 ROIs set in d. From ROIs 2 and 3, which were located at the periphery of the damaged site, non-expanding Ca2+ oscillations were observed over time. (f) A successfully damage-induced ectopic color pattern. This wing is not from the same individual that showed the Ca2+ signals in a-e. Similar results in the same species have been previously published .