Fig. 2

Rgs4 3′-UTR mediates an anterograde transport bias. a Representative phase contrast and GFP fluorescence of hippocampal neuronal culture cotransfected with the MS2+Rgs4 3′-UTR reporter and tdMCP-GFP constructs. Scale bar 20 µm. Asterisk denotes GFP positive cell. b Time series of the dendritic boxed region in a. Representative anterograde (black arrowheads) and retrograde (white arrowheads) moving mRNA granules are indicated. Scale bars 10 µm. c Kymograph of the dendritic region in b. Arrowheads indicate mRNA granules signified in b. Dot plots (d, f, h) and histograms (e, g, i) displaying percentage of anterograde moving granules (d, e), percentage of total anterograde displacement (f, g) and average speed (h, i) for MS2 only or MS2+Rgs4 3′-UTR reporter mRNAs, detected by tdMCP-GFP. In (i), positive values indicate anterograde and negative values indicate retrograde transport. Data represent mean ± standard deviation of three independent experiments (individual experiments shown as gray dots). Asterisks represent p-values obtained by Student’s t test (*p < 0.05, **p < 0.01). Data were obtained from 40-µm dendritic segments at a minimal distance of 20 µm from the cell body. At least 10 dendrites/condition/experiment were analyzed. Total number of dendrites (n_d) and tracks (n_t) analyzed per condition are indicated. Only displacements ≥1.5 µm were considered for analysis