Fig. 4

Overexpression of miR-675 exacerbated muscle atrophy in vivo and in vitro. a Real-time PCR analysis of miR-675-3p expression in gastrocnemius muscles transfected with either miR-675 agomiR or negative control (n = 6). b Representative immunofluorescence images of gastrocnemius muscles transfected with miR-675 agomiR or negative control (n = 6). Left bar panel: cross-sectional area (CSA). Right bar panel: minimal Feret’s diameter (MFD). The anti-laminin stained sections were used for measurement of size of myofibers. Scale bar: 250 μm. c Western blotting and d real-time PCR analyses of Atrogin-1 expression in muscles transfected with miR-675 agomiR or negative control (n = 6). e Real-time PCR analysis of miR-675-3p in C2C12 myotubes transfected with miR-675 mimics or negative control (n = 3). f Representative immunofluorescence images of myotubes transfected with miR-675 mimics or negative control (n = 3). Myotubes were stained with anti-myosin antibody (green). Nuclei were stained with DAPI (blue). Scale bar: 250 μm. g Western blotting and h real-time PCR analyses of Atrogin-1 expression in C2C12 myotubes transfected with miR-675 mimics or negative control (n = 3). The CSA, MFD, the diameters of myotubes, and the intensities of protein bands were quantified using Image-J software. All data are shown as mean ± SD. *P < 0.05 and **P < 0.01 compared to the control groups