Reactive Oxygen Species (ROS) Generation Is Coupled with Cell Delamination
(A) An image of the z-projections of confocal stacks in the pupal notum of a live fly expressing GstD-GFP and Ecad:tdTomato (16 h after puparium formation; APF).
(B) The ratio of delamination in cells positive for GstD-GFP (from 16 to 30 h APF; three nota). Error bar indicates standard error of the mean.
(C) The ratio of GstD-GFP+ cells to delaminating cells (from 16 to 30 h APF; three nota). Error bar indicates standard error of the mean.
(D) A diagram representing the conversion of produced O˙-2 to H2O2 through superoxide dismutase (Sod).
(E) An image of the z-projections of confocal stacks in the pupal notum of a live fly after the injection of a mixture of DHE and Hoechst 33342.
(F) Averaged changes in the ratio of DHE signal to Hoechst 33342 signal (n = 4).
(G) Snapshots from the movie, z-projections of confocal stacks in the pupal notum of a live fly expressing Ecad::GFP (20 h APF). Magenta dots indicate cells that delaminated in 10 h (from 20 to 30 h APF).
(H) The ratio of cell delamination (from 20 to 30 h APF; control: four nota, versus Sod1, Sod2, or Sod3: three nota under the control of Pnr-Gal4). The p value was calculated by one-way analysis of variance (ANOVA) with Dunnett's test. ∗, p < 0.05 and ∗∗, p < 0.01. Error bars indicate standard error of the mean.
(I) Snapshots from the movie, z-projections of confocal stacks in the pupal notum of a live fly expressing SCAT3 under the control of Pnr-Gal4 (16 h APF).
(J) Averaged changes in the FRET ratio (the Venus signal to the ECFP signal) of SCAT3 (16 h APF; control: four nota, versus Sod1: six nota, Sod2: five nota, or Sod3: four nota, under the control of Pnr-Gal4).
(K) Averaged changes in the FRET ratio (the Venus signal to the ECFP signal) of SCAT3 (relative values of the M region to the OM region). The p value was calculated by one-way ANOVA with Dunnett's test. NS, not significant, ∗∗, p < 0.01, ∗∗∗, p < 0.001, and ∗∗∗∗, p < 0.0001. Error bars indicate standard error of the mean.
The boundaries between the midline (M) and outside the midline (OM) regions are outlined with dotted white lines. The anterior-to-posterior axes of all pupae are oriented toward the left. Scale bars: 10 μm. See also Figures S4 and S5.