Fig. 3

Gprc5c^−/− mice show altered ciliary morphology. A Bright-field image of ß-Gal staining of cryosections of Gprc5c^−/− adult mouse OE. ß-Gal staining is confined to the neuronal layer in the adult OE. Dotted line represents basal lamina. B Lower magnification overview image. C Confocal images of adult OE cryosection upon FISH of DIG-labelled probes for Gprc5c (antisense). Gprc5c mRNA was detected in WT, but not in Gprc5c^−/− OE. Dotted line represents basal lamina. Transmission electron micrographs of ultrathin sections OE of WT (D) and Gprc5c^−/− (E). Gprc5c^−/− OE depicts an increased thickness of the ciliary layer. F Scanning electron micrographs of WT OE appears organized with dendritic knobs evenly embedded in the surrounding ciliary network. G Higher magnification. H Gprc5c^−/− OE appears disorganized and has atypical cilia. I Higher magnification. K Cilium with bending of the distal end. L Clumped cilia. M Extracellular vesicles attached to cilia. N Quantification of irregular cilia. Mann–Whitney test showed significant increase in number of irregular cilia in Gprc5c^−/− OE (n = 4, individual data values Additional file 2, error bars represent SEM, *p < 0.05). O Staining of acetylated-tubulin (acTub) as marker for the axoneme of the cilia shows an increase in staining in Gprc5c^−/− OE. P Quantification of staining intensity followed by Student’s t test revealed a significant increase of acetylated-tubulin in Gprc5c^−/− OE (n = 4, individual data values Additional file 2, error bars represent SEM, *p < 0.05). Q Western blot analysis of whole OE preparations, acetylated-tubulin band at 52 kDa, levels were comparable between WT and Gprc5c^−/− OE, equal loading was controlled by actin. Scale bars A, C, O 10 µm; B 500 µm: D, E, F, H 2 μm; G, I 400 nm; K, L, M 200 nm