A dashed oval is drawn for assessment. GAL4 background also generates a small vision phenotype. Similarly, overexpression of PR2 (E) or RNAi mediated knockdown of PR2 (F) in the background does not considerably modify vision size. The dashed oval in panel D has been reproduced in panels E and F to aid in assessment.(TIF) pgen.1002725.s001.tif (2.4M) GUID:?FAF98058-EAEF-4648-9D23-48839B642103 Figure S2: Ack manipulation modifies rpr induced programmed cell death in the wing disc. The TUNEL assay was used to assess the effect of Ack transgenes or alleles on rpr induced programmed cell death. (ACD) TUNEL positive cells are labeled in green and the genotypes are indicated in each panel. (ACD) the TUNEL positive cell images from ACD are superimposed on bright field micrographs of the eye disc. (A) induces programmed cell death in the wing disc. (B) Ack overexpression results in fewer TUNEL positive cells. (CCD) Manifestation of kinase inactive Ack or Ack gene dose reduction shows an increase in TUNEL positive cells.(TIF) pgen.1002725.s002.tif (5.9M) GUID:?2FD0328F-8AB9-4138-AA59-559D1D782CDB Number S3: Akt1 does not modify the hid small vision phenotype. The eye size assay was used to assess the ability of Akt1 loss and gain of function to modify hid induced programmed cell death. Genotypes are indicated in each panel. To aid in vision size comparisons, the dotted oval in panel A is definitely reproduced in panels BCD, the dashed oval in panel E is definitely reproduced in panels FCH and the dashed oval in panel I is definitely reproduced TAK-981 in panels JCL. (ACD) The genetic background combined with (B) Akt1 overexpression, (C) intro of a loss of function allele Akt104226 or (D) overexpression of the constitutively activated mutant Akt1T342D/S505D. (ECH) The genetic background combined with (F) Akt1 overexpression, (G) the loss of the function allele Akt104226 or (H) constitutively triggered Akt1T342D/S505D overexpression. (ICL) Ack overexpression in the genetic background combined with (J) Akt1 overexpression, (K) the loss of the function allele Akt104226 or (L) constitutively triggered Akt1T342D/S505D overexpression.(TIF) pgen.1002725.s003.tif (5.1M) GUID:?CC3910B2-215D-4A96-AEAE-D63B7986CC59 Figure S4: Ack-mCherry and yki-GFP subcellular localization in R-cells. Solitary plane confocal images display TAK-981 the subcellular localization of Ack-mCherry and yki-GFP indicated separately (A and B) or simultaneously (CCE) in third instar R-cells. Nuclei appear as open ringed structures in all TAK-981 panels. (A) Ack-mCherry is definitely nuclear excluded and is found in the cytoplasm and in numerous cytoplasmic puncta. (B) Yki-GFP is also mainly nuclear excluded and more diffusely distributed throughout the cytoplasm compared to Ack. (CCE) Simultaneous manifestation of Ack-mCherry (C) and yki-GFP (D) does not lead to increased nuclear localization of either protein, but does induce yki to co-localize into puncta with Ack. (E) An overlay of panels C and D showing Ack-mCherry (reddish) and yki-GFP (green) localization patterns.(TIF) pgen.1002725.s004.tif (8.9M) GUID:?BE2A0E22-7E2F-48FC-AE83-9E37D307F6C7 Figure S5: Ack expression does not induce transcription of yki targets. Confocal images of third instar vision discs analyzed for manifestation of Ack and beta-galactosidase driven from the ex-lacZ enhancer capture collection ex697 in the absence (ACC) or presence (DCF) of Ack overexpression (posterior is definitely down). The dashed collection indicates the position of the morphogenetic furrow (MF). (A) Beta-galactosidase manifestation pattern in the absence of Ack overexpression shows similar levels of labeling posterior and anterior of the MF. (B) Ack manifestation is definitely higher posterior of the MF. (C) An overlay of panels A and B showing beta-galactosidase (green) and Ack (reddish) manifestation. (D) Beta-galactosidase manifestation pattern in the presence of Ack overexpression again shows similar levels of labeling posterior and anterior of the MF. (E) Overexpression of Ack can be seen posterior to the MF due to GMR driven manifestation. (F) An overlay of panels D and E showing beta-galactosidase (green) and Ack (reddish) TAK-981 manifestation.(TIF) pgen.1002725.s005.tif (6.8M) GUID:?E483576B-B101-4C36-9CF8-9ABB0F865E07 Figure S6: Suppression of hid induced small eye phenotypes by RNAi mediated knockdown of Wwox. The eye size assay was used to assess the ability of Wwox knockdown improve both hid and hidAla5 induced programmed cell death. (A) Hid manifestation inside a and expressing background produces a small vision phenotype. (B) RNAi mediated knockdown of Wwox suppresses the small vision phenotype. A dotted oval is used to aid in comparison. (C) Ack manifestation produces a larger increase in vision size in a similar genetic background. (D) HidAla5 manifestation inside a and expressing background also produces a small vision phenotype. (E) RNAi mediated knockdown of Wwox fails to modify the eye size in the background. A dashed oval is definitely drawn for assessment. (F) Ack manifestation suppresses the small phenotype induced inside Rabbit Polyclonal to Androgen Receptor a background.(TIF) pgen.1002725.s006.tif (2.4M) GUID:?738A01E9-ADD8-4214-82C3-0272E8453340 Abstract Activated Cdc42.