Figure 6. Influence of Vam SH3 Domains on Dachs Localization.
A–B) Horizontal apical sections of wing imaginal discs expressing nub-Gal4 and UAS-Vam-RFP (A,A′) or UAS-Vam-ΔSH3-2:RFP (B,B′). C–E) Horizontal apical sections of wing imaginal discs expressing Dachs:GFP (D:GFP) en-Gal4 UAS-CD8:RFP UAS-dcr2 UAS-Vam-RNAi and also UAS-Vam:RFP (D,D′) or UAS-Vam-ΔSH3-2 (E,E′). Depletion of endogenous Vam and simultaneous expression of Vam-RFP restores D:GFP membrane localization (D,D′), while expression of Vam-ΔSH3-2 (E,E′) fails to fully restore junctional D:GFP. Dashed white line marks approximate edge of en-Gal4 expression. F) Western blot showing results of co-immunoprecipitation by RFP-trap of V5-tagged GFP (negative control) or V5-tagged Ds-ICD (detected by anti-V5), co-expressed in S2 cells with either full length Vam:RFP (Vam-FL) or Vam-RFP lacking the first (Vam-ΔSH3-1) second (Vam-ΔSH3-2) or third SH3 domain (Vam-ΔSH3-1) or containing just the first (Vam-SH3-1) second (Vam-SH3-2) or third SH3 domain (Vam-SH3-3) G). Western blot showing results of co-immunoprecipitation by RFP-trap of V5-tagged GFP (negative control) or V5-tagged Fat-ICD (detected by anti-V5), co-expressed in S2 cells with either full length Vam:RFP (Vam-FL) or Vam-RFP lacking the first (Vam-ΔSH3-1) second (Vam-ΔSH3-2) or third SH3 domain (Vam-ΔSH3-1) or containing just the first (Vam-SH3-1) second (Vam-SH3-2) or third SH3 domain (Vam-SH3-3)(G). See also Fig. S5.