Figure 8. Live imaging of fTRG tagged proteins in living pupal thorax.
(A) Schematic drawing of a 10–12 hr (left) and a 30h pupal thorax (right). The developing epidermis is shown in blue, with the SOP precursors in yellow (developing neurons in red), the differentiating tendons are shown in orange, the myoblasts and muscle fibers in green, and the muscle-tendon junction in red. The schematic positions of the optical sections through epithelium and muscles are indicated with blue and green dotted lines, respectively. (B-Y) Live imaging of pupal thoraces at the indicated stages acquired with a spinning disc confocal (except S and T, which were acquired with a two-photon microscope). Blue bars above the image indicate epithelial sections and green bars indicate muscle sections (as explained in A). Hts-GFP is expressed in fusing myoblasts (B, C) and strongly in developing SOPs (D, E). Dlg1-GFP labels the epithelial junctions (F), internal muscle structures (green dots, G) and an unidentified additional developing epithelium (yellow dots, H, I). Talin-GFP is higher expressed in developing SOPs (J, K) and strongly localised to the muscle-tendon junction from 24 hr APF (red arrowheads, L, M). LanB1-GFP localises to the basal side of the developing epithelium (N) and surrounds the forming muscle fibers (green dots, O-Q) with a slight concentration at the muscle-tendon junction at 30 hr APF (red arrowheads, Q). Act88F-GFP weakly labels the developing epithelium, with a slight concentration in the SOPs until 20 hr APF (R, S) and very strongly marks the IFMs from 24 hr onwards (T, U). βTub60D-GFPis expressed in the fusing myoblasts (V, W) and also labels the microtubule bundles in the developing muscle fibers (X, Y) and hair cells of the developing sensory organs (light blue arrow heads in X). Scale bars indicate 10 µm.
Figure 8—figure supplement 1. Live imaging during pupal development.
