iNeurons as a model to study dynein mediated transport. (A) Example immunofluorescence images of different stages of iNeuron differentiation. DPI 0 = stem cells, DPI 3 = 3 days post induction with doxycycline, DPI 7 = 7 days post induction with doxycycline. (B) Image of 21–23 DPI iNeurons showing staining with axonal (SMI-31, cyan) and dendritic (MAP-2, magenta) markers. Scale bar is 20 µm. (C) Example kymograph of 21–23 DPI Halo-DYNC1H1 treated with 1 nM JFX 554. White arrow highlights processive event. Gray arrow shows diffusive motility. (D) 6 bleaching traces from dynein spots. The traces were separated to enhance clarity. (E) Images of two spots during bleaching from Fig. 2, B and C. The teal spot undergoes 2 bleaching steps and corresponds to the bleaching trace in Fig. 2 B. The light green spot undergoes 6 bleaching steps and corresponds to Fig. 2 C. Scale bar is 0.3 µm. (F) Example kymographs of control (untagged) and Halo-DYNC1H1 21–23 DPI iNeurons treated with 500 nM JFX 554. (G) The intensity of moving dynein spots versus dynein spots treated with NEM (Fig. 2).