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. 2022 Jan 17;11:e73357. doi: 10.7554/eLife.73357

Figure 5. Nord modulates Bone Morphogenetic Protein (BMP) signaling at or above the level of Mad phosphorylation.

(A) Expression of Nord in the wing discs through the third instar larval and early pupal stage. Upper panel: the third instar wing discs from nord-GFP (Mi{PT-GFSTF.2}nordMI06414-GFSTF.2/+) larvae were collected at indicated time points after egg laying (AEL) and were immunostained for GFP (Nord, green) and the pro-domain of Dpp (red). Bottom panel: pupal wings from pupae carrying both nord-GFP were collected at indicated time points after pupation (AP) and were immunostained for GFP (Nord, green) and the pro-domain of Dpp (red). Scale bar, 100 μm. (B) Wing imaginal discs from third instar wild-type and nord mutant (nordMI06414) larvae were immunostained for pMad (green) and Ptc (red). Scale bar, 50 μm. (C) Plotted pixel intensity of pMad or Ptc as a function of A/P position. Each point shows the mean ± SD. n = 15. au, arbitrary units. (D) Anti-pMad staining in wild-type and nordMI06414 pupal wings at indicated hours AP. Scale bar, 100 μm. (E) Quantification of the width (yellow dashed line in panel D) of pMad signal at primordial posterior crossvein (PCV) at indicated time points. Each bar shows the mean ± SD from n > 10 pupal wings. The unpaired two-tailed t-test was used for statistical analysis. ****p<0.0001.

Figure 5.

Figure 5—figure supplement 1. Expression of Nord in the wing discs during the early pupal stage.

Figure 5—figure supplement 1.

(A–C) The early pupal wing discs from nord-GFP (Mi{PT-GFSTF.2}nordMI06414-GFSTF.2/+) pupae were collected at indicated time points after pupation (AP) and were immunostained for GFP (Nord, green) and pMad (red). Note that Nord is expressed in the early pupal wing during the pMad refinement at the posterior crossvein (PCV) primordia. The yellow arrows indicate the PCV primordia. Scale bar, 100 μm.
Figure 5—figure supplement 2. Loss of nord has little effect on the expression of Dpp in wing imaginal discs.

Figure 5—figure supplement 2.

(A) Wing imaginal discs from late third instar wild-type and nord mutant (nordMI06414) larvae were immunostained for the pro-domain of Dpp (green) and Ptc (red). Scale bar, 20 μm. (B) Plotted pixel intensity of Dpp as a function of anteroposterior (A/P) position. The A/P position is determined by the expression of Ptc. Each point shows the mean ± SD. n = 10 wing discs. au, arbitrary units. Scale bar, 50 μm.
Figure 5—figure supplement 3. Venation phenotype in adult wings associated with nord knocking down.

Figure 5—figure supplement 3.

Representative images of the ectopic venation phenotype in adult wings from female flies with the indicated genotypes. Scale bar, 500 μm.
Figure 5—figure supplement 4. Quantification of the ectopic venation phenotype in adult wings associated with nord knocking down.

Figure 5—figure supplement 4.

Quantification of the ectopic venation phenotype in adult wings from female flies with the indicated genotypes. n > 50. Two-sided Fisher’s exact tests were used for statistical analysis. ****p<0.0001, ***p<0.001, **p<0.01, *p<0.05, ns, not significant.