Figure 3.
Cytoplasmic Dynein Was Anchored to the Cell Cortex by Daple in Ependymal Cells
(A) Whole-mount stain of the LV wall with antibodies against dynein intermediate chain (IC, green), tyrosinated tubulin (red), and γ-tubulin or β-catenin (blue). Panels below represent a high magnification of the box shown in the right panel. Arrowheads indicate sites of accumulation of dynein IC with tyrosinated tubulin at the cell cortex.
(B) Lysates of tissue from the LV wall were immunoprecipitated with IgG or anti-Daple antibody. The total cell lysate (Input) and the immunoprecipitates were blotted with anti-dynein IC, anti-dynein HC, anti-Dvl1, or anti-Daple antibodies, as indicated on the left of each panel. Asterisks indicate the bands of co-immunoprecipitated proteins.
(C) Tissue lysates from the LV wall were immunoprecipitated with IgG or anti-dynein IC antibody and immunoblotted with respective antibodies as indicated on the left. Asterisks indicate the bands of proteins that co-immunoprecipitated together.
(D) Whole-mount Daple+/+ and Daple−/− tissue stained with antibodies against dynein IC (green), Daple (red), and γ-tubulin (blue) at the LV wall. Arrowheads indicate sites of co-localization of dynein IC with Daple at the cell cortex.