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. 2020 Apr 20;39(11):e103629. doi: 10.15252/embj.2019103629

Figure EV4. ESCRT and SNARE deficiency in Hh‐receiving versus Hh‐producing cells.

Figure EV4

  1. Clone over‐expressing the Tsp96F‐HA construct, marked with anti‐βgal antibody and immuno‐labelled for endogenous Ptc, white arrowhead indicates the anterior Ptc signal area affected by the clone. Note the slight increase of signal of the Hh target Ptc anterior to the clone (in red rectangle) and compared to a clone‐free area as internal control (blue rectangle). Graph to the right depicts grey values for Ptc signal along the x‐axes, showing the increased signal extension anterior to the clone in red (black arrowhead) compared to the control area in blue.
  2. Basal confocal image of wing disc expressing Hh‐GFP (Bac construct) and immuno‐labelled for endogenous Ptc when expressing the Tetanus toxin (TNT) to block the SNARE complex in Hh‐receiving cells. Arrowhead indicates basal Ptc accumulated in punctuated structures, while yellow arrow outlines the extended distribution length of both the Hh ligand and Ptc signal activation.
  3. Lateral and basal (further right) confocal images of wing discs expressing Hh‐GFP (Bac construct) and immuno‐labelled for endogenous Ptc when expressing the Tetanus toxin (TNT) to block the SNARE complex in Hh‐producing cells. Arrowhead indicates basal Hh accumulation, while yellow arrow outlines the reduced distribution length of both the Hh ligand and Ptc signal activation.
  4. Lateral and basal (further right) confocal images of wing discs expressing Hh‐GFP (Bac construct) and immuno‐labelled for endogenous Ptc in wild‐type conditions. Note shortening of the Ptc gradient and Hh retention in the producing cells when TNT is expressed in Hh‐producing cells versus Ptc gradient extension and greater Hh dispersion when TNT is expressed in Hh‐receiving cells.
Data information: Scale bars 10 μm.