Effects of expression of dpp and tkvQD in a variety of tissues on the timing of pupariation. Effects of expression of dpp and tkvQD in a variety of tissues on the timing of pupariation. (A) Cartoon of larva showing tissues relevant to experiments in (B–G). (B) Expression pattern of genital disc driver AbdB-Gal4 in tissues of 120 h AEL larvae using G-TRACE (RFP, current expression; GFP, past expression). (C) Constitutive expression of dpp at 25°C under control of AbdB-Gal4 causes a developmental delay, whereas tkvQD has no delaying effect. Mean pupariation times are AbdB> 122.2 ± 0.3 h; AbdB>dpp 136.2 ± 2.3 h; and AbdB>tkvQD 122.5 ± 0.9 h. (D) Expression pattern of fat body driver r4-Gal4 in tissues of 120 h AEL larvae using G-TRACE (RFP, current expression; GFP, past expression). (E) Constitutive expression of dpp at 25°C under control of r4-Gal4 causes a severe delay. Although larvae reach the L3 stage, they remain in L3 and do not pupariate for the duration of the experiment while tkvQD has no delaying effect. Mean pupariation times are r4> 128.0 ± 0.5 h and r4>tkvQD 127.3 ± 0.8 h (n = 500 for r4>dpp represents the number of L1 larvae that were raised under density control.). (F) Expression pattern of dpp-LG in tissues of 120 h AEL larvae using lexOp-mCD8::GFP (GFP, current expression). (G) Constitutive expression of lexOEGFP::dpp at 25°C under control of dpp-LG causes a delay compared with the average pupariation timing of the individual dpp-LG and lexOEGFP:dpp transgenes. Mean pupariation times are dpp-LG> 118.3 ± 1.5 h; lexOEGFP::dpp 125.2 ± 1.0 h; and dpp-LG>lexOEGFP::dpp 148.2 ± 1.3 h. Linda Setiawan et al. LSA 2018;1:e201800216 © 2018 Hariharan et al.