Figure 6: PCC plays a role in the rotational direction of genitalia.
(a) The angle between the cell boundary (dashed line), labelled for Discs Large (Dlg; white) and the AP axis (yellow arrow) was defined as θ (blue and red arcs). Scale bar, 10 μm. (b) Frequency of cell boundaries with an angle θ of −90° to 0° or of 0° to 90° to the AP axis in controls, myoID, DE-Cad knockdown and sqh knockdown flies at 23 and 29 h APF. (c) Rose diagrams indicating the frequency of external genitalia orientation in adult male DE-Cad knockdown flies. White arrow indicates the direction from the analia to the external genitalia. (d) First image in a time-lapse series showing genitalia rotation. Cells were assigned different numbers and colours, and their rightward velocity with respect to the AP axis was tracked (yellow arrow). Rightward movement was defined as positive unidirectional motion (white arrow). A time-lapse series for d is shown in d’. Scale bar, 50 μm. (e) Velocity of rightward cellular movement with respect to the AP axis. (f) Frequency of cell boundaries forming an angle θ with the AP axis at each time point, in the control flies analysed in d. (g) Gene expression levels for myoID and myoIC were analysed by qRT–PCR. Green bars are GFP-positive cells and white bars are GFP-negative cells in the dissected tissue. *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001. Error bars indicate s.d. (b,e,f,g). Number of flies, N; number of cell boundaries, nboundary (b,e and f); number of experimental replicates, n (g). Fly genotypes: UAS-Dcr2/+; AbdB-GAL4 UAS-RedStinger/+ (a,b, Control), myo31DFk2 (b, myoID), UAS-Dcr2/UAS-shg dsRNA; AbdB-GAL4 UAS-RedStinger/+ (b,c, DE-Cad dsRNA), UAS-Dcr2/UAS-sqh dsRNA; AbdB-GAL4 UAS-RedStinger/+ (b, sqh dsRNA) Nrg::GFPG305/Y (d–f). NS, not significant.
