Fig. 2

Differentiation advantage of ciliated neurons as compared to non-ciliated neurons in a LUHMES WT background. A The percentage of ciliation in differentiating human LUHMES neurons increases once neurons exit the cell cycle (d1), peaks in differentiating neurons (d3) and then decreases during the later stages of neuron differentiation and maturation (d4-d6). The percentage of neurons reaching stage 3 of differentiation steadily increases throughout the entire differentiation and maturation process (n = 134–394). B-E Populations of ciliated LUHMES neurons (n = 39–128) are more efficient than non-ciliated LUHMES neurons (n = 15–116) in breaking the bipolar symmetry of stages 2a and 2b, and thus, in promoting axon outgrowth (d1-d3 summarized and individual days). F Identification by immunocytochemistry of nuclei (Hoechst staining), centrosomes/basal bodies (PCNT marker), cilia (ARL13B marker) and axons (TAU marker) reveals that the majority of stage 3 differentiating neurons in a given population are ciliated. Arrowheads point to stage 3 neurons either being ciliated (in green) or non-ciliated (in red). Mean values ± s.e.m. are shown. The results are from a minimum of three independent experiments with a total of at least six technical replicates. We conducted regular two-way ANOVA analyses (not repeated measures) with multiple comparisons (Bonferroni’s test) between groups. *p < 0.05; **p < 0.005; ***p < 0.0005; ****p < 0.0001