Fig. 5. Localization of dynein 2 and LIC3 in the cilia. (A-F) Double labeled confocal images of photoreceptor cells. The connecting cilia are stained with K26 mAb (red, Cy3); counter-stained (green, Alexa 488) with rabbit anti-dynein 2 HC (A,D,E) Ab, anti LIC3 Ab (B,F), or anti-acetylated -tubulin mAb (C). Bars, 10 µm (A,F). Arrows in A and B indicate the connecting cilia. Asterisks in B indicate LIC3 staining in the external limiting membrane, the zone of junctional complexes between adjacent photoreceptors and glial cells; the nature of this staining is unknown. (A,B,D-F) Most cells show a concentration of both dynein 2 HC (A,D,E) and LIC3 (B,F) on the proximal side of the connecting cilia (large arrows); occasionally also on the distal side (D,F, small arrows). (C) Acetylated -tubulin staining shows extension of axoneme from the basal body (large arrows) through the connecting cilium to the distal side (small arrows). The yellow to orange color in the connecting cilium represents overlap of the two antigens. (G-N) Dynein 2 HC within primary cilia in cultured cells. NRK cells are stained with rabbit polyclonal antisera (G,I,K,M); counterstained with mAbs (H,J,L,N), respectively. Primary cilia (arrows in A-L) are stained with anti-detyrosinated tubulin Ab (Glu-Tub; G), anti-acetylated -tubulin Ab (Ac-tub; H,J,L), also with anti-dynein 2 HC Ab (D2 HC; I) and anti-LIC3 Ab (K). Cytoplasmic staining of dynein 2 HC (M) was not colocalized with Golgi 58K marker protein (N). Bar, 10 µm (H).

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