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. Author manuscript; available in PMC: 2017 Oct 24.
Published in final edited form as: Dev Cell. 2016 Oct 13;39(2):254–266. doi: 10.1016/j.devcel.2016.09.017

Figure 5. Vam and Dachs Reciprocally Regulate each other and Physically Interact.

Figure 5

A) Wing imaginal disc expressing Dachs:GFP (D:GFP, green) in clones and UAS-vam-RNAi in posterior cells (right side) under en-Gal4, which leads to loss of junctional D:GFP. Right panels show close-ups of the anterior (1) and posterior (2) boxed areas. (B,B′) Horizontal (above) and vertical (below) sections of a wing disc expressing en-Gal4 UAS-Vam:RFP (red); D:GFP (green) showing increased junctional D:GFP in posterior cells. (C, C′) Horizontal (above) and vertical (below) sections of a wing disc expressing en-Gal4 UAS-dcr2 UAS-d-RNAi; Vam:GFP loss of membrane localization of Vam:GFP (green) in posterior cells. (D,D′) Horizontal (above) and vertical (below) sections of a wing disc expressing en-Gal4 UAS-D:V5 (red); Vam:GFP (green) showing increased levels of membrane localized Vam:GFP in posterior cells. (E) Western blot showing results of co-immunoprecipitation experiments on proteins co-expressed in S2 cells. Schematic depicts the different Dachs fragments used with the amino acid numbers below. V5-tagged GFP (negative control), Full length Dachs (D:V5), D N terminus(D–N:V5), D Middle domain (D-M:V5) or D C terminus (D-C:V5) were co-expressed with full length Vam:RFP, and complexes were precipitated using RFP-trap. (F) Western blot showing results of co-immunoprecipitation experiments on proteins co-expressed in S2 cells. Schematic depicts Vam and the SH3 domain fragments used with the amino acid numbers below. V5-tagged GFP (negative control) or Full length Dachs (D:V5) were co-expressed 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-ΔSH3-3), and complexes were precipitated using RFP-trap. GAPDH was used as a control for loading and transfer in the input lanes. See also Fig. S4.

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