Primary cilia are hair-like sensory organelles protruding from the surface of most human cells. As cilia are dynamic, several aspects of their biology can only be revealed by real-time analysis in living cells. Here we describe the generation of primary cilia reporter cell lines. Furthermore, we provide a detailed protocol of how to use the reporter cell lines for live-cell imaging microscopy analysis of primary cilia to study their growth as well as intraciliary transport. For complete details on the use and execution of this protocol, please refer to Bernatik et al. (2020) and Pejskova et al. (2020).

Charles University Faculty of Science Albertov 6 128 00 Prague Czechia

Department of Experimental Biology Faculty of Science Masaryk University Kamenice 5 62500 Brno Czechia

Laboratory of Adaptive Immunity Institute of Molecular Genetics of the Czech Academy of Sciences Vídeňská 1083 14220 Prague Czechia

Laboratory of Cell Motility Institute of Molecular Genetics of the Czech Academy of Sciences Vídeňská 1083 14220 Prague Czechia

Laboratory of Cilia and Centrosome Biology Department of Histology and Embryology Faculty of Medicine Masaryk University Kamenice 3 62500 Brno Czechia

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Tau-tubulin kinase 2 restrains microtubule-depolymerizer KIF2A to support primary cilia growth

BBSome-deficient cells activate intraciliary CDC42 to trigger actin-dependent ciliary ectocytosis