Fig 6. Ecdysone signaling and Br cooperate to induce differentiation.

Flip-out clones, GAL4 expression, and MARCM clones are marked with GFP and outlined in yellow. (A) Anti-Cut staining (magenta) is absent during late L3 when br is knocked down in the posterior compartment using en-Gal4. (B) cut (magenta) is ectopically expressed in Flip-out clone cells misexpressing br-Z1 during mid L3. (C) cut (magenta) is not expressed in MARCM clones mutant for both chinmo and br (chinmo¹;br^RNAi cells) during late L3 (11/12 clones, n = 7 discs). (D) br-Z1 (magenta) is silenced in Flip-out clones expressing both EcR^DN and chinmo^RNAi transgenes in late L3 (52/52 clones, n = 4 discs). (E) cut (magenta) and sens (blue) are not expressed in Flip-out clones expressing both EcR^DN and chinmo^RNAi transgenes during late L3 (15/15 clones, n = 7 discs). (F) cut (magenta) is not expressed in Flip-out clones expressing both br-Z1 and EcR^DN transgenes during late L3 (4/4, n = 3 discs). (G) Schematic outline of the above experiments. Scale bars: 30 μm. br, broad; CW, critical weight; EcR^DN, dominant negative form of ecdysone receptor; eL3, early L3; en, engrailed; FO, Flip-out; GFP, green fluorescent protein; L3, third larval stage; MARCM, Mosaic Analysis with a Repressible Cell Marker; RNAi, RNA interference; sens, senseless.