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. 2018 Jan 24;6(1):52–64.e4. doi: 10.1016/j.cels.2017.10.015

Figure 5.

Figure 5

A Ligand Induction-Specific Network of Wnt Signaling

(A) Correlation-based network connecting components of Wnt secretion and receptor binding with potential co-regulatory modules. “Bait” genes required for ligand secretion and receptor binding (green nodes) were selected and potentially novel components (light-blue nodes) were called if they shared a CSI > 0.9 in the Wnt-active state. Potential negative regulators (orange nodes) were added if they shared a between-state similarity (PCC > 0.5) with the destruction complex (red nodes) and a Wnt-active state-specific dissimilarity (PCC < −0.65) with at least one “bait” gene. The network was further extended for genes sharing a CSI > 0.9 with potentially novel positive or negative regulators (gray nodes). Genes were connected if their state-specific profiles shared an absolute PCC > 0.65 or if their between-state shared an absolute PCC > 0.5 (dashed lines). Positive and negative correlations are shown as blue and red lines, respectively. Per state, the modules of positive regulators (green, light blue), negative regulators (orange), and the destruction complex (red) were separately arranged using a force-directed layout algorithm. The edge width represents the absolute PCC.

(B) State-specific (if |PCC| > 0.65) and between-state (if |PCC| > 0.5) relations of Catsup with the ligand secretion machinery and negative regulators.

(C) Catsup single depletion effect on Wnt signaling activity upon increasing Wg concentrations.

(D and E) Knockdown of Catsup in third instar wing imaginal disc leads to Wg accumulation in the Wg-producing cells. Total Wg staining intensity (red) was quantified using FIJI (n > 6 wing discs). enGal4, UAS-GFP/UAS-Catsup RNAi, posterior to the right (GFP positive). Scale bars, 20 μm.

(F and G) Knockdown of Catsup leads to depletion of extracellular Wg. Extracellular Wg (Wg ex) was stained on living third instar imaginal disc without cell permeabilization. Staining intensity (red) was quantified using FIJI (n > 6 wing discs). enGal4, UAS-GFP/UAS-Catsup RNAi, posterior to the right (GFP positive). Scale bars, 20 μm.