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. 2019 Jul 10;8:e44752. doi: 10.7554/eLife.44752

Figure 10. Formation of RNA clusters at protrusions is promoted by translational inhibition and requires microtubules.

PolyA RNA was detected in NIH/3T3 cells with the indicated treatments. Boxed regions are enlarged to show the presence (arrows) or absence of polyA RNA granules at the tips of protrusions. Graph shows scoring of protrusions for the presence of polyA RNA granules. Values are mean and standard error of at least three independent experiments. For each experiment approximately 300 protrusions from more than 25 cells were observed. p-value: *<0.02, **<0.01, ***<0.001 by one-way ANOVA with Dunnett’s multiple comparisons test, compared to control. Scale bars: 10 μm.

Figure 10.

Figure 10—figure supplement 1. PolyA RNA staining is a more reliable identifier of peripheral clusters in 3T3 cells.

Figure 10—figure supplement 1.

Enlarged views of individual protrusions from cells processed by in situ hybridization to detect the Pkp4 RNA or polyadenylated RNA. Single Pkp4 RNAs can be observed at protrusions in association with accumulated polyA RNA signal indicative of peripheral clusters (upper panels; white arrows). We also observe peripheral polyA RNA clusters that do not contain any detectable Pkp4 RNAs (lower panels; yellow arrowhead). Scale bars: 5 μm (upper panels), 10 μm (lower panels).