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. 2013 Sep 11;8(9):e74598. doi: 10.1371/journal.pone.0074598

Figure 4. Interactions between poly(A)+ mRNA bound to MTRIP lacking flag tag and cytoskeletal elements in human dermal fibroblasts (HDF).

Figure 4

(A) β-tubulin, vimentin and phalloidin IF, poly(A)+ mRNA, and PLA between poly(A)+ mRNA, and the cytoskeletal elements in HDF were imaged with a laser-scanning confocal microscope. Merged images of the cytoskeleton (white), poly(A)+ mRNA (red), PLA (green), and nuclei (blue) are shown. Single image plane is represented. Scale bar, 10 µm. (B) The mean MTRIP volume was similar (Kruskal-Wallis One Way ANOVA on Ranks, p=0.08) in cells, where the interactions between β-actin mRNA and β-tubulin (n=18, mean=404µm3, s.d.=317µm3), vimentin (n=19, mean=518µm3, s.d.=362µm3), or phalloidin (n=15, mean=653µm3, s.d.=211µm3) were quantified. (C) The mean percentage of MTRIP colocalized with PLA (PLA-MTRIP) was similarly minimal (Kruskal-Wallis One Way ANOVA on Ranks, p=0.15) in β-tubulin (n=18, mean=0.01%, s.d.=0.03%), vimentin (n=19, mean=0.04%, s.d.=0.1%), or phalloidin (n=15, mean=0.20%, s.d.=0.41%). (D) The mean PLA frequency was also minimal (Kruskal-Wallis One Way ANOVA on Ranks, p=0.23) in β-tubulin (n=18, mean=0.0004µm-3, s.d.=0.001µm-3), vimentin (n=19, mean=0.0008µm-3, s.d.=0.0001µm-3), or phalloidin (n=15, mean=0.001µm-3, s.d.=0.002µm-3). Error bars, s.d.