DEV027789 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure S1 -

  Fig S1. Notch protein levels and expression of the Notch target gene E(spl)-mβ during wing development. (A-D) Wild-type (A) and omb-gal4, UAS-GFP/UAS-N^dsRNA (B-D) late third instar wing discs labelled to visualize Notch protein (red or white; A,B,D), Wingless (Wg, pink; C) and GFP (B-D) expression. Note, in B-D, reduced expression levels of Notch or Wg proteins in the N^dsRNA-expressing cells (green). (E-J) E(spl)-mβ-lacZ expression visualized by histochemical staining for β-gal activity in wild-type (E,G,I), sd-gal4, UAS-N^dsRNA (F) and omb-gal4, UAS-N^dsRNA (H,J) wing discs of the following larval stages: early second (E,F), early third (G,H) and late third (I,J) instar. d, dorsal; v, ventral compartments.

• Supplemental Figure S2 -

  Fig S2. Expression of the sd-Gal4 driver in the presence or absence of Notch activity during wing development. (A-H) sd-gal4, UAS-GFP (A,C,E,G), sd-gal4, UAS-GFP/UAS-Ser^TM (B) and sd-gal4/UAS-N^dsRNA, UAS-GFP (D,F,H) wing discs of the following larval stages: early second (A,B), late second (C,D), early third (E,F) and late third (G,H) instar, labeled to visualize GFP (green) expression. Note, no change in GFP expression in early discs, while the GFP expression domain shows a mirror image duplication pattern, probably due to the duplication of notum structures (nt and nt').

• Supplemental Figure S3 -

  Fig S3. Tissue growth induced by overexpressed CycE or Dcdc25 (String). Histogram showing the size (in arbitrary units) of wild type, sd-Gal4, UAS-CycE and sd-Gal4, UAS-Dcdc25/String early second instar wing discs raised under the same conditions. The average wing disc sizes and standard deviations were 1±0.22 (wild type), 1.7±0.4 (sd>CycE) and 1.13±0.22 (sd>stg). sd>CycE and sd>Stg wing discs were significantly bigger than wild-type discs (P<10⁻¹³ and P=0.01, respectively). The number of scored discs was 39 (sd>CycE), 33 (sd>Stg) or 47 (wild type).

• Supplemental Figure S4 -

  Fig S4. Notch activity is not affected by cell cycle progression. (A-E) ptc-gal4 UAS-GFP (A), wild type (B), ptc-gal4 UAS-GFP, UAS-N^intra (C), ptc-gal4 UAS-GFP, UAS-N^intra UAS-CycE (D), and ptc-gal4 UAS-GFP, UAS-N^intra UAS-String (E) late third instar wing discs labelled to visualize Wg (red, B-E) and GFP (green, A,C-E) protein expression. Endogenous levels of Wg along the dorsoventral (dv) boundary are shown in B. Note similar levels of ectopic expression of Wg in the ptc domain (labelled by GFP) in C-E. (F-H) C96-gal4 UAS-GFP UAS-Ser^TM (F), C96-gal4 UAS-String UAS-Ser^TM (G) and C96-gal4 UAS-CycE UAS-Ser^TM (H) late third instar wing discs labelled to visualize Cut (red) and GFP (green) protein expression. Note similar levels of repression of Cut expression in F-H.

• Supplemental Figure S5 -

  Fig S5. Early DV compartmentalization defects in the absence of Wg or Notch activity. (A-I) apterous-lacZ (ap-lacZ) expression visualized by histochemical staining for β-gal activity in third instar wing discs of the following genotypes: wild type (A), sd-Gal4; UAS-Ser^TM (B), sd-Gal4; UAS-N^dsRNA (C), sd-Gal4; UAS-notum (D), sd-Gal4; UAS-notum, UAS-N^iINTRA (E), sd-Gal4; UAS-notum, UAS-CycE (F), sd-Gal4; UAS-Ser^TM, UAS-Wg (G), sd-Gal4, UAS-Ser^TM, UAS-CycE (H) and sd-Gal4, UAS-Dl^TM, UAS-CycE (I). apterous expression is restricted to the dorsal (d) compartment (A). Note failure to restrict apterous to the most dorsal part in B-F. Note rescue of this phenotype in G-I. d, dorsal; v, ventral compartments.

• Supplemental Figure S6 -

  Fig S6. Coupling growth and DV compartmentalization. (a,B) Illustration of the subdivision of the wing imaginal disc into dorsal (blue) and ventral (white) compartments by the signaling molecules Wingless (Wg, orange) and Vein (Vn, green). (A) Dorsally expressed Vn induces the expression of the dorsal selector gene apterous through activation of the EGF receptor pathway (EGF-R*). Wg represses Vn expression restricting it to the dorsal region. (B) Growth promoted by the activity of Notch restricts EGF receptor pathway activity and apterous expression to the most dorsal part of the wing primordium.