Fig. 6
Comparative analysis of FRET vs DRET signals in stage-10A egg chambers. (A–F) Confocal images of stage-10A egg chambers hybridized with FRET (A–C) and DRET (D–F) pairs. FRET signals were collected after excitation at 514 nm while DRET signals were collected after excitation at 458 nm. (A–C) Signals recovered in the presence of the acceptor only AP-Cy5 (A), the donor-only DP-Cy3 (B) or the combination of both probes (C). A significant basal signal is observed in follicular cells and the posterior part of the oocyte. (D–F) Signals recovered in the presence of the acceptor only A0p (D), the donor-only Dp (E) or the combination of both probes (F). For enhanced visualization, grey levels are color-coded using the Fire LUT. (G) Distributions of signal-to-background intensity ratios. Average intensities for the FRET (ex. 514 nm, em. 650–700 nm, shown in blue) and DRET signals (ex. 458 nm, em. 650–700 nm, shown in red) were normalized for each egg chambers to the average intensities of the posterior follicular cells. In total 30 egg chambers were analyzed across three distinct experiments (represented as distinct color shades) for both FRET and DRET pairs. The white horizontal lines indicate the average values of the results. Scale bar shown in (A) represents 20 µm. ****P < 0.0001 (Mann–Whitney test).
