Fig. 8

Monitoring local translation in living neurons. a A diagram depicting the preparation of rat primary cortical neurons for imaging. b The dendrite of a sample living neuron expressing frankenbody (FB-GFP) and the smHA-KDM5B-MS2 translation reporter (n = 15 cells in two independent experiments). White arrows indicate translation sites that were tracked, as depicted in the cartoon on the left. The spatiotemporal evolution of one mRNA track with directed motion is shown through time. c Puromycin treatment of a representative cell. Two circled translation sites were tracked as they disappeared following the addition of puromycin. Upper-right: The mean number of nascent chain spots normalized to pre-puromycin levels decreases after puromycin treatment (N = 48 spots from three cells in three independent experiments). Error bars, cell-to-cell SEM. d The travel distance through time for the translation spot highlighted in b. Gray highlights in d and black arrows in b indicate directed motion events. e Plot of velocities faster than 1 µm/s (37 velocities from eight cells in two independent experiments). Mean: 1.40 ± 0.07 mm/s (spot-to-spot SEM). See also Supplementary Fig. 6. Scale bars, 10 µm. Source data are provided as a Source Data file. For the box and whisker plots, median is shown by a white line, the box indicates 25–75% range, and whiskers indicate 5–95% range