Localisation of Nek8 in the cilia of Lewis and LPK. Immunohistochemical labeling for acetylated α-tubulin, γ-tubulin and Nek8 in LPK ( A, B, C) and Lewis ( D, E, F) animals. Single labeling for acetylated α-tubulin (panels A and C) confirmed the abnormal length of cilia in the LPK animals. Double labeling with Nek8 (red) and γ-tubulin (green, as a marker for the basal body) indicated that Nek8 was in close proximity to the base of the cilia in the Lewis ( E), but was located at variable positions along the cilia in the LPK ( B). Double labeling with Nek8 (red) and acetylated α-tubulin (green, panels C and F) confirmed the altered expression of Nek8 along the length of the cilia in the LPK ( C). Arrows highlight examples of cilia identified by either acetylated α-tubulin or γ-tubulin. Scale bars in each panel represent 5μm.