Feline Lindhout

3 Centrosome-mediated microtubule remodeling during axon formation in human iPSC-derived neurons 103 Supplementary Figure 2. Centriole loss perturbs axonal Trim46 targeting and action potential maturation A. Typical examples of human iPSC-derived NSCs/neurons (day 3) with 0 (control) or 3 days Centrinone-B treatment, and immunostained for Ki67, β3-Tubulin and MAP2. Scale bar: 15 µm in overview. B. Quantifications of the ratio MAP2-positive/Ki67-positive Centrinone-B treated or control human iPSC-derived NSCs/neurons at day 1, 3 and 5. N=2, n=54-110 cells. C. Quantifications of the ratio β3-Tubulin-positive/Ki67-positive Centrinone-B treated or control human iPSC-derived NSCs/neurons at day 1, 3 and 5. N=2, n=49-104 cells. D. Quantifications of the percentage of MAP-positive Centrinone-B treated or control human iP- SC-derived neurons at day 1, 3 and 5. N=2, n=121-165 cells. E. Quantifications of the percentage β3-Tubulin-positive Centrinone-B treated or control human iPSC-derived neurons at day 1, 3 and 5. N=2, n=121-165 cells. F. Representative examples of evoked AP firing in Centrinone-B treated human iPSC-derived neu- rons recorded at day 11. Shown is the response to a single depolarizing current step of a neuron that fires no APs, a neuron that fires a single AP and a neuron that fires multiple APs. The offset current (I hold ) was adjusted to keep the baseline membrane potential at approximately −60 mV (dashed lines). G. AP amplitude recorded in Centrinone-B treated and control human iPSC-derived neurons of 10-11 days (control: n=15 cells, +Centrinone-B: n=14 cells) and 13-14 days (control: n=36 cells, +Centrinone-B: n=27 cells). H. AP half-width recorded in Centrinone-B treated and control human iPSC-derived neurons of 10-11 days (control: n=15 cells, +Centrinone-B: n=14 cells) and 13-14 days (control: n=36 cells, +Centrinone-B: n=27 cells). I. Resting membrane potential of Centrinone-B treated (n=54 cells) and control (n=64 cells) human iPSC-derived neurons. J. Input resistant of Centrinone-B treated (n=54 cells) and control (n=63 cells) human iPSC-derived neurons. Data information: Data represent mean ± SEM. Chi-square-test including post-hoc analysis with Bonferroni correction ( D, E ); Student’s t test ( F, G : control 10-11 vs 13-14 days); Mann Whitney test ( G : control vs Centrinone-B, I ). *** p<0.001, ** p<0.01, * p < 0.05, ns p>0.05