Fig. 4

Leg primordia and newly added legs in the late preparatory and rigidation periods. A-F Autofluorescence images obtained by confocal laser scanning microscope of transparentized samples of stadium V juveniles at 405 (cyan), 488 (green), 561 (red), and 640 nm (blue). A,B Ventral view of the posterior several rings. C,D Higher magnification of the white boxed region in A and B, respectively. E,F Lateral view of the posterior several rings. A,C,E Leg primordia at the late preparatory period. B,D,F Transparent protrusions at the rigidation period. G-J SEM images with the covered cuticles removed. G,H Stadium V juveniles. I,J Stadium VI juveniles. G,I Ventral view of two pairs of leg primordia at the late preparatory period. Especially, leg primordia of G are of ring 16. H,J Lateral view of two pairs of newly added legs at the rigidation period. In SEM images, red and blue colorations, respectively, indicate anterior and posterior leg pairs. In C-H, each leg was labeled L1–8 so that the corresponding leg could be identified. Anterior to the left in all images. Scale bars show 100 μm (A-F) and 50 μm (G–J). cx, coxa; fe, femur; pf, prefemur; po, postfemur; ta, tarsus; ti, tibia