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. 2012 Nov 13;7(11):e49105. doi: 10.1371/journal.pone.0049105

Figure 1. Cell Ablation Strategy.

Figure 1

(A) Strategy used to produce Ablating and Non-Ablating larvae. w*; P{Sgs3-GFP}3 females were crossed to w*;+;rn-GAL4,UAS-egr,tubGAL80ts/TM6Tb, tubGAL80 males to give rise to the Ablating genotype (w*;+;rn-GAL4,UAS-egr,tubGAL80ts/Sgs3GFP) and the control Non-Ablating genotype (w*;+;TM6Tb, tubGAL80/Sgs3GFP). (B) Strategy to induce cell ablation. Embryos of the Ablating and Non-Ablating genotypes were collected at room temperature in four hour intervals and transferred to 18°C. First-instar larvae (48 hours AEL) were transferred to a vial containing standard cornmeal-yeast-agar medium and were allowed to develop at 18°C until the designated time for ablation induction. At the designated time during L3 (130–230 hours AEL) vials were transferred to 30°C for 40 hours, returned to 18°C and monitored daily to document the time to Sgs3GFP expression, pupariation or eclosion. RNA for qPCR and samples for EIA experiments were collected at time points T0–T3.