DEV032672 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure S1 -

  Fig. S1. Critical weight determines the developmental response to starvation on 2% agar in w¹¹¹⁸ larvae. (A,B) Wing imaginal discs from larvae at 5 hours (h) after third instar ecdysis (AL3E). (C,D) Discs from larvae starved on 2% agar from 5-30 hours AL3E. (E,F) Discs from larvae fed on standard food until 30 hours AL3E. (G,H) Wing discs from larvae at 15 hours AL3E. (I,J) Discs from larvae starved on 2% agar from 15-39 hours AL3E. (K,L) Wing discs from larvae staged to 39 hours AL3E. Shown is expression of (A,C,E,G,I,K) Wingless (WG) and (B,D,F,H,J,L) Senseless (SENS). Asterisks indicate the sensory organ precursors (SOPs) of the wing hinge chordotonal organ (F,J,L). The SOPs of the triple row at the wing margin are marked by double arrowheads (F,J,L). The arrowheads in F, J and L point to bristle SOPs in the wing pouch and notum. Scale bar: 100 µm.

• Supplemental Figure S2 -

  Fig. S2. The expression of anterior/posterior axis genes does not change in response to nutritional conditions in w¹¹¹⁸ larvae. The expression of Patched (A-F) and Engrailed (G-L) was examined in pre-critical weight and post-critical weight larvae under different nutritional conditions. (A,G) Wing discs from larvae staged to 5 hours (h) after third instar ecdysis (AL3E). (B,H) Discs from larvae fed on sucrose alone between 5 and 29 hours AL3E. (C,I) Discs from 29 hour AL3E larvae. (D,J) Discs from 15 hour AL3E larvae. (E,K) Discs from larvae fed on sucrose alone between 15 and 39 hours AL3E. (F,L) Discs from 39 hour AL3E larvae. Scale bar: 100 µm.

• Supplemental Figure S3 -

  Fig. S3. The expression of housekeeping genes does not change in response to nutrition in w¹¹¹⁸ larvae. The expression patterns of Armadillo (A-F), Actin as determined by phalloidin stain (G-L) and Tubulin (M-R) were examined in pre-critical weight and post-critical weight larvae under different nutritional conditions. (A,G,M) Wing discs from larvae staged to 5 hours (h) after third instar ecdysis (AL3E). (B,H,N) Discs from larvae fed on sucrose alone pre-critical weight (between 5 and 29 hours AL3E). (C,I,O) Discs from 29 hour AL3E larvae. (D,J,P) Discs from 15 hour AL3E larvae. (E,K,Q) Discs from post-critical weight larvae fed on sucrose alone (between 15 and 39 hours AL3E). (F,L,R) Discs from 39 hour AL3E larvae. Scale bar: 100 µm.

• Supplemental Figure S4 -

  Fig. S4. Enhancing insulin signaling in the prothoracic gland, using the phantom GAL4 driver, changes the developmental response of wing discs to starvation. (A-F) Wing imaginal discs from phantom (phm)>GFP larvae. (G-L) Discs from phm>Dp110 larvae. (A,B,G,H) Wing discs from larvae staged to 5 hours (h) after third instar ecdysis (AL3E). (C,D,I,J) Discs from larvae fed on sucrose alone between 5 and 29 hours AL3E. (E,F,K,L) Discs from larvae at 29 hours AL3E. Shown is expression of (A,C,E,G,I,K) Wingless (WG) and (B,D,F,H,J,L) Senseless (SENS). The asterisks in F, J and L are the sensory organ precursors (SOPs) of the wing hinge chordotonal organ. The double arrowheads in J and L mark the SOPs in the triple row at the wing margin. Other SOPs in the wing pouch and notum are marked by arrowheads (F,J,L). Scale bar: 100 µm.

• Supplemental Figure S5 -

  Fig. S5. Suppressing insulin signaling in the prothoracic gland represses the differentiation of Wingless and Senseless in larvae fed on sucrose alone post-critical weight. The expression of WG (A,C,E,G,I,K) and SENS (B,D,F,H,J,L) in wing imaginal discs from P0206>GFP larvae (A-F) and P0206>PTEN larvae (G-L). Wing discs are from (A,B,G,H) larvae at 15 hours after third instar ecdysis (h AL3E), (C,D,I,J) larvae fed on sucrose alone from 15-39 hours AL3E, or (E,F,K,L) from larvae at 39 hours AL3E. The double arrowheads in D and F mark the triple row of bristle sensory organ precursors (SOPs). The asterisk marks the wing hinge chordotonal organ SOPs and the arrowheads denote the bristle SOPs in the wing pouch and notum in D and F. Scale bar: 100 µm.

• Supplemental Figure S6 -

  Fig. S6. Suppressing EcR expression in wing discs causes premature differentiation of the Cut expression pattern in pre-critical weight larvae fed on sucrose alone. The expression of Cut in wing imaginal discs at 5 hours (h) after third instar ecdysis (AL3E) (A,D), in discs from larvae fed on sucrose alone from 5-29 hours AL3E (B,E) and at 29 hours AL3E (C,F). (A-C) C765 larvae; (D-F) C765>EcRi larvae. Scale bar: 100 µm.

• Supplemental Figure S7 -

  Fig. S7. Overexpressing the isoforms of Broad is insufficient to drive premature differentiation in the wing discs of pre-critical weight larvae fed on sucrose alone. The expression of Wingless (WG) (A,C,E,G,I,K,M,O,Q,S,U,W,Y,AA,CC) and Senseless (SENS) (B,D,F,H,J,L,N,P,R,T,V,X,Z,BB,DD) in wing imaginal discs from C765 (A,B,K,L,U,V), C765>Broad Z1 (C,D,M,N,W,X), C765>Broad Z2 (E,F,O,P,Y,Z), C765>Broad Z3 (G,H,Q,R,AA,BB) and C765>Broad Z4 (I,J,S,T,CC,DD) larvae. (A-J) Discs at 5 hours (h) after third instar ecdysis (AL3E). (K-T) Discs from larvae fed on sucrose alone from 5-29 hours AL3E. (U-DD) Discs from larvae staged to 29 hours AL3E. The arrowheads in V, X, Z and BB mark bristle sensory organ precursors (SOPs) in the wing pouch and notum. The SOPs of the triple row are shown with a double arrowhead in V, X, Z and BB. The asterisks in V, X, Z, BB and DD indicate the SOPs of the wing hinge chordotonal organ. C765>BR Z4 larvae had misshaped wing discs that lacked SENS in the SOPs of the hinge chordotonal organ and several other bristle precursors in the notum and wing pouch. Scale bar: 100 µm.