RBF<sup>D253A</sup> is pro-apoptotic in the ZNC and induces more apoptosis than RBF in third instar larvae wing imaginal discs.

<p>(A, E) <i>C96-Gal4</i> and <i>vg-Gal4</i> expression patterns are visualized by <i>UAS-mtGFP</i> expression in third instar larvae wing imaginal discs. (B-D, F-H) Apoptotic cells are labeled by TUNEL in wing imaginal discs; specific staining of apoptotic cells corresponds to bright white patches. (B, F) <i>C96-Gal4/+</i> and <i>vg-Gal4/+</i> control discs have few apoptotic cells. (C) <i>C96-Gal4/UAS-RBF</i> wing discs are similar to control. (D) Some apoptotic cells are observed within the <i>C96-Gal4</i> expression domain in <i>UAS-RBF<sup>D253A</sup>/X</i>; <i>C96-Gal4/+</i> discs (white arrow). (G, H) Apoptotic cells are observed within the <i>vg-Gal4</i> expression domain in <i>vg-Gal4/+</i>; <i>UAS-RBF/+</i> and <i>UAS-RBF<sup>D253A</sup>/X</i>; <i>vg-Gal4/+</i> wing discs (white arrows). (G) The white arrowhead indicates a zone at the center of the pouch where cells are not TUNEL-labeled in <i>vg-Gal4/+</i>; <i>UAS-RBF/+</i> wing discs. All discs are shown with posterior to the top.</p>