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. 2017 Oct 1;144(19):3612–3624. doi: 10.1242/dev.152736

Fig. 5.

Fig. 5.

Hh requires Disp to modulate cytonemes. (A,A′) S2 cells expressing cholesterol-modified Hh (A, magenta) or cholesterol-free HhN (A′, magenta) were fixed using MEM-fix, immunostained and imaged by confocal fluorescent microscopy. Phalloidin (green) marks actin. DAPI (blue) marks nucleus. Arrows indicate HhN present in the cytoneme. Scale bar: 5 µm. (B) Cytonemes were quantified in S2 cells expressing the indicated proteins. Cytoneme occurrence was quantified in ∼75 cells per condition over three independent experiments and all data pooled. Error bars indicate s.e.m. Significance was determined using a one-way ANOVA test (***P≤0.001). (C) Endogenous disp was knocked down in control, cytoplasmic mCherry- or Hh-expressing S2 cells. Kinesin serves as loading control. Disp is detected as a broad band (bracket). A non-specific band (ns) recognized by the antibody is indicated. (D) S2 cells were transfected with Hh or cytoplasmic mCherry expression vectors in the presence of control or disp3′UTR dsRNA. Cytoneme occurrence was determined as in B. Error bars indicate s.e.m. Significance was determined using a one-way ANOVA (**P≤0.01).