Statistical evaluation as described in Figure 1D: nubbin. GFP, nubbin. Lig. Genotypes: w/y w; ey Gal4/UAS GFP w/yw, ey Gal4/, UAS ligR185C/ yw hsFLP/yw, UAS GFP/, Act. CD2. Gal4, UAS GFP/ yw hsFLP/yw, Act. CD2. Gal4, UAS GFP/UAS lig yw/Y; nubbin Gal4/UAS GFP yw/Y; nubbin Gal4/, UAS lig/. Figure S3 Lig will not co localize with DART1, and endoge nous Lig, FMR1 and Capr co localize with Rin Cherry. S2 cells co transfected with GFP lig, RFP FMR1, RFP rin, RFP DART1, and GFP FMR1, RFP rin and HA ligR185C. S2 cells stained with DAPI to visualize DNA and with a HA to visualize HA Lig. S2 cells co transfected with GFP ligR185C and RFP DART1 don’t reveal any co localization. S2 cells have been stained with DAPI to visualize DNA. Scale bar represents 25 mm. Untransfected S2 cells stained for endogenous Ago1. S2 cells had been stained with DAPI to visualize DNA.
Scale bar represents 25 mm. S2 cells transiently transfected with GrinCherry to express Rin Cherry at endogenous levels. In many of the cells Rin Cherry is homoge neously in the cytoplasm of transfected cells. In few cells Rin selleck chemicals Cherry types punctae and localizes with Lig, FMR1 and Capr. S2 cells have been stained with DAPI to visualize DNA. Scale bar represents 25 mm. Untransfected S2 cells stained for endogenous Lig and Capr. Lig and Capr localize in bigger punctae but not in cells with smaller punctae. S2 cells have been stained with DAPI to visualize DNA. Scale bar represents 25 mm. Figure S4 Lig and Rin fragments display no autoactivity in Y2H experiments. Adverse controls for Y2H interactions among Lig, LigFG LA, Rin, Rin1 175, Rin129 492 along with the empty vector.
Lig, Rin, Rin1 175, Rin129 492 and LigFG LA fused towards the AD and towards the DBD, respectively, usually do not show autoactivity. Figure S5 Analysis of rin hypomorphic alleles and also the genomic rescue transgene GrinCherry. Barasertib Aurora Kinase inhibitor Negatively marked 72 h old rin2, PGawBrinNP3248 and PGawBrinNP5420 mutant clones in eye imaginal discs of third instar larvae. Rin Cherry levels expressed from the GrinCherry are autonomously increased within the rin mutant clones. The scale bar represents 50 mm. Scanning electron micrographs of adult PGawBrinNP3248 and PGaw BrinNP5420 eyes generated by eyFLP/FRT mediated mitotic recombination. The scale bar represents one hundred mm. Statistical analyses as described in Figure 1D: handle, PGawBrinNP3248 and PGawBrinNP5420. The extended slender pupae formed by FMR1D113M/D50M rinNP3248/2 and FMR1D113M/D50M rinNP5420/2 are rescued with 1 copy on the GrinCherry transgene.
The controls usually do not show any defects. Statistical analysis on the rescue of FMR1D113M rin2 mutant eyes with Grin as described in Figure 1D: handle, FMR1D113M rin2 Grin; FMR1D113M rin2.