Stability of TS51D, TS51D2xSce and TS58A2xSce transgene vectors at DSBs induction under different heat shock delivery as indicated by absence or presence of marker phenotypes.

All vectors contain two recognition sites for homing endonucleases. DSBs can be induced in TS51D sequentially (columns I-SceI and I-CreI) or simultaneously (I-SceI + I-CeI). Two breaks are induced in TS51D2xSce and TS58A2xSce. For optimal induction of I-SceI strong heatshock is necessary, and for I-CreI – light one. Figures for full one-step transgene stabilization are in bold. Statistical analysis is provided in Supplementary Table S1.a(G+) GFP expression in these phenotypes is masked by RFP expression.bsummary stabilization after deletion of at least one ITR (sum of W-(G+)R+, W+G+R- and W-G+R-).cDSB induction by I-CreI endonuclease was carried out in flies with already stabilized transgene (without 3′ITR and white).