Characterization of Xenopus Deup1 antibody and time course analyzis of MCCs markers in A6-MCI cells. (A) Cos1 cells were transfected with the indicated construct and immunoblotted with anti-GFP (left panel) or homemade anti-Deup1 antibody (right panel). (B) A6, A6-MCI, or St18 Xenopus animal caps injected with the indicated construct, or morpholinos targeting Deup1, were immunoblotted with anti-Deup1 antibody. The Deup1-specific band disappears in animal caps injected with 3 distinct morpholinos against Deup1. (C) Western blot showing the expression of Deup1, centrin, and Ac-α-Tub in A6-MCI cells at the indicated times after induction. (D) Confocal pictures of A6-MCI cells stained with Deup1, centrin, and ZO1 antibodies reveal A6-MCI differentiation stages. At the stage of centriole production (left), Deup1 and centrin reveal globular structures, which correspond to deuterosomes surrounded by growing centrioles. Later (middle), centrin staining reveals released centrioles associated with Deup1 puncta. In mature cells (right), Deup1 is no longer detectable at apically positioned centrioles. Source data are available for this figure: SourceData FS2. Original Ponceau membrane and gel of data shown in Fig. S1. Ponceau-stained membranes were cut along indicated dotted lines. Membranes were incubated with the indicated antibodies. Red rectangles indicate the regions of the gel that are shown in Fig. S1. Note that colorimetric and chemiluminescent ladders were loaded in the same well in Fig. S2 B.