1. Vps36 interacts with Smo in the absence of Hh
Vps36 interacts with Smo in the absence of Hh. (A–E) S2 cells were transfected with the indicated constructs, treated with or without Hh, and followed by Co-IP with anti-Myc antibody and western blot for anti-Myc to detect wild-type Smo or Smo mutation expression, or for anti-V5 to detect wild-type Vps22, Vps25, Vps36, Vps36GLUE expression in the Co-IP products or cell lysates, respectively.
Vps36 interacts with Smo in the absence of Hh. (A–E) S2 cells were transfected with the indicated constructs, treated with or without Hh, and followed by Co-IP with anti-Myc antibody and western blot for anti-Myc to detect wild-type Smo or Smo mutation expression, or for anti-V5 to detect wild-type Vps22, Vps25, Vps36, Vps36GLUE expression in the Co-IP products or cell lysates, respectively. Arrows indicate the bands where Vps36–V5 or Vps36GLUE–V5 interacted with Myc–SmoWT or Myc-tagged Smo mutations. (A) In ESCRT-II complex, only Vps36 interacted with wild-type Smo. Vps36 only interacts with Smo in the absence of Hh (B). This interaction is regulated by Smo C-tail phosphorylation (C) and the phosphorylation clusters (D). (E) GLUE domain of Vps36 (Vps36GLUE–V5) interacts with Smo in the absence of Hh.
Xiaofeng Yang et al. J Cell Sci 2013;126:
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