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Fig. 2. Endogenous Inv is exclusively detected at the base of primary cilia (the Inv compartment) but not in the basal body or ciliary necklace segment. (A) Localization of endogenous Inv. Inv was detected with the anti-Inv antibody in wild-type and inv cells. Primary cilia were stained with the anti-acetylated 
-tubulin antibody (ac-tubulin; green), and the basal body or centrosome were stained with the anti-
-tubulin antibody (green). The Inv signal (red) is detected at the base of the primary cilium, but not in the basal body area. In inv cells, there is no Inv signal in primary cilia. Thus, the antibody recognizes true endogenous Inv. Scale bars: 10 µm. Line scans of the fluorescent signal from `*' to `**' of the inserted images are shown on the right. A.U., arbitrary units. (B) Immunoelectron microscopical images of wild-type cells probed with an anti-Inv antibody. Significant Inv staining can be seen at the proximal segment of the primary cilium (Inv compartment), but not at the distal segment. The Inv signal is only seen between the ciliary membrane and axoneme, and not in the ciliary necklace segment (CN), basal body (BB) or transition zone (TZ). (C) Immunoblotting with anti-Inv antibody. Cell lysates extracted from inv cells (lane –) and inv cells expressing Inv protein (lane +) were separated by 7.5% SDS-PAGE, transferred to PVDF membrane and stained with anti-Inv antibody. A single major band is detected at approximately the predicted Inv molecular weight [117 kDa in lane + (arrowhead)].
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