Figure 4.

Wdr78 is required for ciliary assembly of Dynein-f. (A) Experimental scheme of ependymal cilia purification for LFQ mass spectrometric analysis. (B and C) Examination of purified cilia by immunofluorescence staining (B) and negative-staining electron microscopy (C). AC-tub- and Hydin-labeled axonemes and the CP, respectively. (D) Proteome analysis of a cilia preparation from Ctrl-i-treated mEPCs. Only hits with ≥5 unique peptides were included. (E) Relative LFQ intensities of the indicated proteins in 78-i2-treated cilia preparation, normalized to corresponding Ctrl-i-treated sample. (F) Dnah2 was markedly reduced in Wdr78-depleted ependymal cilia. AC-tub and Ift52 served as positive controls for the cilia preparations, whereas Lamin B1, a nuclear lamina protein, and GM130, a Golgi protein, served as negative controls. We failed to detect Wdr63 in the ciliary samples due to insufficient protein concentration.