Figure 4. Hiw and Wnd Regulate Dscam Expression through Dscam 3'UTR.

(A–D) The Dscam 3’UTR is required by hiw to regulate Dscam expression in C4da neurons. Dscam[TM2] ∷ GFP transgenes with or without UTRs (schematically shown at the top) were co-expressed with mCD8 ∷ mRFP in wild-type (wt) or hiw^ΔN C4da neurons using the C4da driver Gal4^4–77. Quantification of the immunofluorescence intensities is shown at the bottom. Sample numbers are shown in the bars. (E) Wnd promotes Dscam expression through the Dscam 3’UTR in C4da neurons. Dscam[TM2] ∷ GFP transgene containing 3’UTR, was co-expressed with either Wnd (OE Wnd) or the membrane protein rCD2 (OE CD2) as a control, together with mCD8 ∷ mRFP, by the C4da driver Gal4^4–77. In A–E, Dscam[TM2] ∷ GFP immunofluorescence in ddaC cell bodies were normalized to that of mCD8 ∷ mRFP and presented as % of controls. Scale bars: 5 mm. (F) Wnd promotes Dscam expression through the Dscam 3’UTR in cultured S2 cells. Dscam constructs were transfected into S2 cells along with either an empty vector (control) or a Wnd-expression construct (Wnd). Dscam ∷ GFP expression was examined using Western blotting with an anti-GFP antibody. The intensities of Dscam ∷ GFP bands were normalized to those of tubulin and presented as fold change for statistical analysis (n = 4)