Fig 3. For and Gyc76C are required for the refinement and maintenance of BMP signaling.

(A,B) PCV regions of 24 hour AP wild type (wt) wings showing anti-pMad staining (A) and suppression of anti-DSRF expression (B) in veins. (C,C’) 24 hour AP for ⁰² homozygote wing showing loss of loss of pMad (C) and failure to suppress DSRF (C’) in PCV. (D,D) 23 hour AP hh-Gal4 UAS-gyc76C-RNAi wings showing loss of pMad (D) but still slight suppression of DSRF (E) in PCV. (F) 28 hour AP hh-Gal4 UAS-gyc76C-RNAi wing showing failure to suppress DSRF in PCV. (G) Anti-pMad staining in 20 hour AP wild type wing. (H) 20 hour AP en-Gal4 UAS-gyc76C-RNAi wing showing abnormally broad anti-pMad in PCV and LVs. (I) 20 hour AP for ⁰² homozygous wing showing pMad in PCV. (J-K’) Anti-pMad staining (red, white) in homozygous gyc76C ^3L043 (J,J’) or gyc76C ^KG0372ex33 (K,K’) clones (identified by absence of green GFP) in 28 hour AP hs-FLP/+; gyc76C FRT ^2A /hs-GFP RpS17 ⁴ FRT ^2A wings. Individual cells in PCV retain high levels of pMad (arrows), similar to levels in neighboring wild type PCV or LV cells. (L,L’) Normal anti-pMad staining (red, white) in PCV cells of homozygous for ⁰² clone (identified by absence of green RFP) in hsFlp; for ⁰² FRT ^40A/ubi-RFP FRT ^40A 26 hour AP wing. (M) Anti-pMad (red) and anti-Myc (green) staining in a 24 hour AP UAS-myc-gyc76C/+; hh-Gal4/+ wing. Arrow indicates ectopic pMad between L3 and L4 anterior to the PCV, outside the region of hh-Gal4-driven expression of Myc-Gyc76C. (N,O) Comparison of anti-pMad (red) staining in 25 hour +/for ⁰² and for ⁰²/for ⁰² wings with hh-Gal4 UAS-gyc76C-myc/+ (anti-Myc, green). Ectopic pMad observed in for heterozygote (N) is lost in for homozygote (O).