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. 2017 May 25;8:15201. doi: 10.1038/ncomms15201

Figure 8. miR-29b is sufficient to induce muscle atrophy in vivo.

Figure 8

(a) qRT–PCR analysis showed increased miR-29b, but not miR-29a or miR-29c expressions, in mice treated with miR-29b agomir compared to negative control (NC agomir) (n=5 per group). (b) miR-29b agomir induced muscle atrophy, as determined by gastrocnemius muscle morphology, gastrocnemius weight (GW) and gastrocnemius weight/body weight (GW/BW) ratio (n=5 per group, scale bar, 1 cm). (c) The grip strength of right hind limb was reduced in miR-29b agomir-treated mice (n=5 per group). (d) miR-29b agomir-induced muscle atrophy was also evidenced by haematoxylin–eosin (HE) staining, periodic acid-schiff (PAS) staining and succinate dehydrogenase (SDH) staining (n=5 per group, scale bar, 50 μm). (e) Quantification of muscle fibre diameter distribution confirmed that miR-29b agomir induced muscle atrophy (n=5 per group). (f) Quantification of diameter of different myofibre types showed that all types of fibres underwent atrophy in miR-29b agomir-treated mice (n=5 per group). (g) qRT–PCR analysis showed decreased Myh1, Myh2, Myh4 and Myh7 expressions in mice treated with miR-29b agomir (n=5 per group). (h) qRT–PCR analysis showed upregulated Atrogin-1 and Murf-1 expressions in mice treated with miR-29b agomir (n=5 per group). (i) Western blot analysis showed upregulation of Ubiquitin protein expressions in mice treated with miR-29b agomir (n=3 per group). (j) qRT–PCR analysis showed downregulated MHC in mice treated with miR-29b agomir (n=5 per group). (k) Western blot analysis showed reduced MHC protein level in mice treated with miR-29b agomir (n=3 per group). (l) Western blot analysis showed downregulated P62 but up-regulated LC3-II protein levels in mice treated with miR-29b agomir (n=3 per group). (m) qRT–PCR analysis showed that mtDNA copy number was decreased in miR-29b agomir-treated mice (n=5 per group). Age- and sex-matched mice were used for experiments randomly. Error bars, s.e.m. An unpaired, two-tailed Student's t-test was used for comparisons between two groups. *P<0.05, **P<0.01.