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Fig. 5 | Skeletal Muscle

Fig. 5

From: Fragile X mental retardation protein regulates skeletal muscle stem cell activity by regulating the stability of Myf5 mRNA

Fig. 5

FMRP binds Myf5 transcripts and affects rates of deadenylation. a Illustration of Myf5 transcripts (Mus musculus) with FMRP binding WGGA (UGGA, AGGA) consensus sites indicated. Consensus sites which are within close proximity to each other and form potential FMRP binding G-quadraplex sites (gray) are indicated. One such G-quadraplex is in proximity to the microRNA-31 (miR-31) binding site on the 3’UTR of Myf5. b Immunoprecipitation (IP) of FMRP-mRNA complexes from C2C12 cells transfected with plasmids encoding GFP and FLAG-hFMRP. Immunoprecipitating antibodies are against FLAG-hFMRP (anti-FLAG, left) and GFP (anti-GFP, right). Amplification of Myf5 (upper panels) and Pax7 (lower panels) transcripts by semi-quantitative RT-PCR are shown, with thermocycle numbers indicated. c Poly(A) tail lengths of Myf5 (left) and Pax7 (right) transcripts amplified from total RNA isolated from 3-day cultures of satellite cells isolated from muscle of adult Pax3 GFP/+ (wild-type, wt) and Fmr1 −/− mice. Estimated polyA lengths are indicated adjacent to each acrylamide gel. Gene-specific primers (GSP) against Myf5 and Pax7, with product size indicated, are shown in the lower panels. A red dotted line indicates Myf5 polyA lengths < 200 nucleotides present in wild-type but not in Fmr1 −/− MuSCs

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