Nub gal4

For most experiments Drosophila strains were cultured in standard cornmeal-agar medium (#789211, NutriFly BF, Genesee Scientific) and maintained at 25 °C. The RNAi experiments were performed at 28 °C as previously described [35 (link)]. The sau^z2217 mutant strain was described previously [35 (link), 67 (link)]. UAS-dGOLPH3 RNAi (#46150, [35 (link)]) and UAS-Tctp RNAi (# 26632) were obtained from the Vienna Drosophila Resource Center Collection (VDRC, [44 (link)]). The following fly strains were from the Bloomington Drosophila Stock Center (BDSC, Indiana University, Bloomington, IN, USA): UAS-Rheb (#9689), UAS-GFP (#4775), Df(2 L)Exel7010 (#7782), en-Gal4 (#30564), nub-Gal4 (#86108), ey-Gal4 (#5534), elav-Gal4 (#8765), and bam-Gal4 (#80579, [68 (link)]). The following fly strains were obtained from FlyORF (University of Zurich, [69 (link)]): UAS-dGOLPH3-HA (#F002769), UAS-Tctp-HA (#F002479), UAS-14-3-3ζ-HA (#F001064). bam-Gal4 and tub-Gal4/tub-Gal80^ts (gift of Dr. Timothy Megraw, Florida State University, USA) were used to deplete dGOLPH3 respectively in spermatocytes and in larval/pupal tissues and to express dGOLPH3-HA, Tctp-HA, and 14-3-3ζ-HA proteins. The fly strains expressing either GFP, or fluorescent tagged-dGOLPH3 were described previously: GFP [34 ], dGOLPH3-RFP [34 ], GFP-dGOLPH3 [35 (link)], and GFP-dGOLPH3^K167A/R170L [35 (link)].

Tefu(dATM)^RNAi (V22502), mei-41(dATR)^RNAi (V103624), grapes(dChk1)^RNAi (V12680), Loki (dChk2)^RNAi (V110342), dp53^RNAi (V38235) and basket(dJNK)^RNAi (V104569) lines were obtained from the Vienna Drosophila Resource Center (VDRC), and UAS-p53^H159N (BL8420) line from the Bloomington Stock Center. his1^RNAi (31617R-3) line was obtained from NIG-FLY and is described in ref. ^{11 (link)}. GFP^RNAi and hp1a^RNAi lines are described in ref. ^{61 (link)}. nub-GAL4 and UAS-Dic2 were kindly provided by Dr. Casali and are described in Bloomington Stock Center. UAS-puc^2A was kindly provided by Dr Martín-Blanco. UAS-RNH1 (human) line was prepared by subcloning the corresponding cDNA from pcDNA3-RNH1^{63 (link)} into pUAST, and injected for random integration into w¹¹¹⁸ embryos.
dH1 depletion was induced in the pouch region of the wing imaginal discs and in polytene chromosomes using his1^RNAi; nub-GAL4; UAS-Dic2 flies by themselves or combined with either additional RNAi lines or overexpression constructs. Similarly, HP1a depletion was induced in the same tissues using hp1a^RNAi; nub-GAL; UAS-Dic2 flies. To analyze wing phenotypes, wings were prepared and recorded as described^{11 (link)}. Wing images were measured and analyzed using FIJI software⁶⁴ .

y w was used as a wild-type strain. The following mutants and transgenic lines were used: nos-gal4VP16[14] (link) from R. Lehmann (New York University, New York, NY, USA); nub-gal4 and ap-gal4[15] (link) from T. Hayashi (NIG, Mishima, Japan); UAS-H2B-ECFP[16] (link) from S. Kondo (NIG, Mishima, Japan); UAS-EgfrDN[17] (link) from M. Freeman (MRC Laboratory of Molecular Biology, Cambridge, UK); UAS-pav-GFP[18] (link) from Y. M. Yamashita (University of Michigan, Ann Arbor, MI, USA); bam-GFP[19] (link) from D. McKearin (HHMI, Chevy Chase, MD, USA); and argos^delta7[20] (link) from the Bloomington Stock Center (Indiana University, Bloomington, IN, USA). goe^5–11 and goe³³¹ alleles were generated by imprecise excision of a P element, EY01697, inserted in the 5′UTR of goe.