Feline Lindhout

Quantitative mapping of transcriptome and proteome dynamics during polarization of human iPSC-derived neurons 2 65 Supplementary Figure 1. Successful and protracted transition of developmental stages in human iPSC-derived neurons A . Representative images of neurite morphology at different locations (axon, dendrite) at day 7 and 13. Scale bar: 5 µm. B. Quantifications of neurite width at different locations (axon, dendrite) at day 7 and 13. N=3, n=20- 23. C. Representative image of a polarized human iPSC-derived neuron (day 13) immunostained for Trim46, PanNaV and AnkG. Zoom represents the AIS structure. Scale bars: 10 µm in overview, 5 µm in zooms. D. Number of APs versus input current injection of hiPSC-derived neurons (day 7-14) that fire multiple APs (red, n=20 cells) or fire only one AP (blue, n=32 cells). We excluded two cells in the multiple firing group, due to variation in baseline. Shown is the mean ± SEM. E. Resting membrane potential of human iPSC-derived neurons that fire a single AP (n=32 cells) or multiple APs (n = 22 cells). F. Input resistance of human iPSC-derived neurons that fire a single AP (n=32 cells) or multiple APs (n=21 cells). G . AP after-hyperpolarization of human iPSC-derived neurons that fire a single AP (n=32 cells) or multiple APs (n=22 cells). H. Maximum sodium current of human iPSC-derived neurons that fire a single AP (n=22 cells) or multiple APs (n=17 cells). Used tests: One-way ANOVA with Tukey’s post-hoc analysis ( B ), Mann-Whitney U test ( E - H ); *** p<0.001, ** p<0.01, * p<0.05, ns p≥0.05; error bars are ± SEM.