Adhesive micropatterns facilitate the study of centrosome migration during primary ciliogenesis. (A) Primary ciliogenesis is a multistep process that is proposed to begin with centrosome maturation and the formation of a ciliary vesicle at the distal end of the mother centriole, after which the centrosome migrates to the apical surface and attaches to the cortex. Full extension of the axoneme occurs once the mother centriole is anchored to the cortex. (B) Side view of micropatterned RPE1 cells expressing EGFP-centrin1 (white), cultured in the presence or absence of serum for 24 h, and stained with phalloidin to visualize F-actin, acetylated tubulin antibody to label the cilium, and DAPI to stain the DNA. (C) Measurement of centrosome z position as a percentage of nuclear height. Migration started within 2 h of serum starvation and appeared completed 6 h later. Measurement of the proportion of ciliated cells showed a delayed process compared with centrosome migration (one experiment, n = 60 cells per condition). Error bars represent standard deviation. (D) Side view of a representative time-lapse imaging of serum-starved RPE1 EGFP-centrin1 cells on micropatterns. Centrosome migration was engaged 1 h after starvation and completed 2 h later. (E) Representation of time-lapse centrosome movement in serum-starved RPE1 EGFP-centrin1 (data of three independent experiments, n = 53 cells). (i) The graph represents all the raw data. (ii) The centrosome trajectories are synchronized, i.e., they start when the centrosome leaves the basal pole. (iii) The plotting of the centrosome trajectories is limited to the maximal z position, i.e., they come to an end when centrosomes reach the apical pole. (F) Frequency distribution of centrosome migration velocity from basal pole to apical pole. Bars, 5 µm.