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. 2018 Oct 17;145(20):dev165712. doi: 10.1242/dev.165712

Fig. 4.

Fig. 4.

Temporal differences in response to elevated Wts or reduction of cell density. (A,B) Wing discs expressing en-Gal4 UAS-GFP tub-Gal80ts UAS-Wts stained for DNA (blue) and ex-lacZ (red). Larvae were cultured at 18°C, shifted to the restrictive temperature for Gal80ts (29°C) for 24 h before dissection and dissected at ages corresponding to 84 h (A) and 120 h (B) AEL. (C) Histogram showing quantitation of relative posterior to anterior (P/A) ex-lacZ intensity for the wing discs at 84 h and 120 h AEL with Wts overexpression. n=14 at 84 h and 12 at 120 h. (D) Wing disc expressing en-Gal4 UAS-dcr2 tub-Gal80ts UAS-Cdk1-RNAi stained for Dcr2 (blue) and E-cad (red). Larvae were cultured at 18°C, shifted to the restrictive temperature for Gal80ts (29°C) for 24 h before dissection at an age corresponding to 108 h AEL. The rightmost panel is a cell area color map (blue to violet, 0 µm2 to 12 µm2). (E) Average recoil velocities after laser cutting of cell junctions in anterior (A) or posterior (P) compartments of wing discs from control animals, or those expressing cdk1 RNAi or Mmp2 in posterior cells. n=17 (control), 19 (cdk1 RNAi) or 19 (UAS-Mmp2). (F,G) Wing discs expressing en-Gal4 UAS-GFP UAS-dcr2 tub-Gal80ts UAS-Cdk1-RNAi ex-lacZ stained for expression of ex-lacZ (red) and DNA (blue). Larvae were cultured at 18°C, shifted to the restrictive temperature for Gal80ts (29°C) for 24 h (F) or 36 h (G) before dissection. (I) Wing disc expressing en-Gal4 UAS-dcr2 tub-Gal80ts UAS-Cdk1-RNAi stained for DNA (blue) and Yki (green). Larvae were cultured at 18°C, shifted to the restrictive temperature for Gal80ts (29°C) for 24 h before dissection. (K,M) Wing discs expressing en-Gal4 UAS-dcr2 tub-Gal80ts UAS-Cdk1-RNAi Sqh:GFP (K) or Jub:GFP (M) stained for Dcr2 (blue) and E-cad (red). Larvae were cultured at 18°C and shifted to the restrictive temperature for Gal80ts (29°C) for 24 h before dissection. Pictures on the right show higher magnification of boxed area in central panel. (H,J,L,N) Histograms showing relative posterior to anterior (P/A) ex-lacZ (H), nuclear Yki (J), Sqh:GFP (L) and Jub:GFP (N) intensity, normalized to DNA (H,J) or E-cad (L,N) for wing discs of the indicated genotypes. n=12 (ex-lacZ, Sqh:GFP and Jub:GFP) or 22 (Yki). Error bars indicate 95% confidence interval. ns, P>0.05; **P≤0.01; ***P≤0.001; ****P≤0.0001. Scale bars: 20 µm.