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. 2011 Mar 7;192(5):797–811. doi: 10.1083/jcb.201007014

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© 2011 Laursen et al.

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Figure 2. — Integrin activation specifically enhances translation of mRNA containing the MBP 3′UTR. (A) Oli-neu cells cotransfected with pcDNA-MBP or pcDNA-MBP-3′UTR in combination with pEGFP-C1, wild-type β1-integrin, or the β1(D759R) and stained for EGFP or β1-integrin (green) and MBP (red) as indicated. Note the increased expression of MBP from the pcDNA-MBP-3′UTR construct only in cells expressing the D759R mutant. (B) Quantification of the experiment shown in A. For each experiment at least 100 cells were analyzed and the mean percentage of EGFP+ or β1+ cells expressing MBP were plotted ±SD. Statistical significance was analyzed by Students t test. **, P < 0.01; ***, P < 0.001. Note that coexpression with β1(D759R) reverses the inhibitory effect of the 3′UTR of MBP mRNA on MBP expression. (C) Analysis by flow cytometry of EGFP expression in Oli-neu cells triple transfected with pEGFP or pEGFP-MBP-3′UTR in combination with an empty vector (pcDNA3.1), wild-type β1-integrin, or β1(D759R) and a DsRed-expressing vector as a control for transfection efficiency (see Fig. S1). Note that β1(D759R) specifically enhances the translation of the EGFP variant containing the 3′UTR of MBP mRNA, as shown in the right plot by the shift to the right of the black histogram relative to the green histogram (that shows the control level of EGFP expression as seen in the cells transfected with the pEGFP). (D) The translation index calculated from the experiment in C as described in Materials and methods, with values being mean ratios from three independent experiments ±SD. Statistical significance was analyzed by one-way ANOVA followed by a Tukey’s multiple comparison test. **, P < 0.01. Note the significant increase in translation only in cells expressing the activated integrin.

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