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. 2012 May 15;31(13):2908–2921. doi: 10.1038/emboj.2012.130

Figure 5.

Figure 5

RhoG expression is regulated by miR-124-binding sites in the 3′UTR of the RhoG mRNA. (A) Scheme of the RhoG message including the miR-124-binding sites in the 3′UTR. (B) Diagram of the sensor construct. (C) Immunoblotting for EGFP expression showed the importance of the miR-124-binding sites for sensor expression. Immunoblotting for β-tubulin served as a loading control. (D) Scheme for experimental design (upper panel). (E) The EGFP expression in hippocampal neurons for the indicated constructs is shown. (F) The EGFP expression of the sensor construct EGFP–3′UTRfull(RhoG) was downregulated by endogenous miR-124. Point mutations in the miR-124-binding sites (see Materials and methods) reduced the degree of regulation. Mean values (n=22 neurons)±s.e.m. (**P<0.005; *P<0.05). Scale bar, 20 μm. (G) Quantification of relative fluorescence intensities of cell bodies of hippocampal neurons transfected with the indicated constructs at DIV8 and stained for endogenous RhoG at DIV8+2. Mean values (n=21 neurons for EGFP+anti-miR-control, and n=18 neurons for EGFP+anti-miR-124)±s.e.m. (**P<0.005). (H, I) Anti-miR-124 inhibited the expression regulation of the sensor construct. Mean values (n=76 neurons)±s.e.m. (***P<0.001). Scale bar, 20 μm.