Eyeless gal4

The following lines were acquired from the Bloomington Stock Center: w¹¹¹⁸ (BL# 38690), hsFlp,hsIscel/CyO (BL # 6934), nanos-Gal4 (BL# 32179), Tubulin-Gal4 (BL# 5138), eyeless-Gal4 (BL# 8227), In(1)white^m4 (BL# 807), Sb[V] (BL# 878), Cre (BL# 1501) lid¹⁰⁴²⁴ (BL# 12367), lid^k06801 (BL# 10403). PSR^FM1 was provided by Kristin White (Massachusetts General Hospital, Charlestown, MA), FRT40A, UTX^{1 (link)} was provided by Andreas Bergmann (M. D. Anderson Cancer Center, Houston, TX), FRT40A, UTX^Δ was provided by Jürg Müller (MPI of Biochemistry, Chromatin and Chromosome Biology, Martinsried, Germany).

Fly husbandry was conducted according to standard procedures. All crosses were performed at 25 or 27 °C. The WT strain used was Oregon R. Transgenic flies for expression of UAS-Polo-GFP (WT and mutants) were created by site-directed insertions after injections of our plasmids (pUAS-K10attB-based) in the attP154 strain by BestGene. Insertions on the third chromosome allowing very similar expression levels were selected for experiments (PhiC31 integrase-mediated site-specific transgenesis, BestGene Inc). The UAS-Polo RNAi strain used for depletion of Polo was obtained from Vienna Drosophila Resource Center (#20177). Expression of Polo transgenes in the early embryo was driven by matα4-GAL-VP16 (#7062, Bloomington Drosophila Stock Center). Expression of Polo transgenes and depletion of endogenous Polo in the developing head tissues was driven by eyeless-Gal4 (#5534, Bloomington Drosophila Stock Center).
For fertility tests, well-fed females were mixed with males in tubes containing grape juice agar and allowed to lay eggs for 1 day before being removed. The percentage of hatched embryos was counted 24 h later.
For viability tests, flies were crossed and the number of observed flies (number of pupae hatching) relative to their expected number in the progeny (total number of pupae) was expressed as a percentage.