FIGURE 7.

TCONS-00036665 overexpression inhibits mouse skeletal muscle development in vivo. (A) The injection scheme of lenti-TCONS-00036665 or lenti-EV particles into the hindlimb muscles of C57 mice. The injection was performed every 1 week, and the mice were sacrificed after a month injection. (B) qPCR results showing that TCONS-00036665 expression was significantly increased. (C) Western blotting results showing that the protein expression of MyoD, MyoG, and MyHC was significantly inhibited by TCONS-00036665 overexpression, and the relative MyoD, MyoG, and MyHC protein expression was quantified by ImageJ software. (D) Representative pictures of quadriceps (Qu), tibialis anterior (TA), and gastrocnemius (Gas) muscles from the right or left hindlimbs of mice. (E) The statistical analysis of the weight of Qu, TA, and Gas muscles from the right or left hindlimbs of nine injected mice; the results showed that overexpression of TCONS-00036665 decreased the weight of each muscle. (F,G) Representative pictures of myosin (F) and dystrophin (G) immunofluorescence staining of the cross section of Qu, TA, and Gas muscles in mice showing that TCONS-00036665 overexpression remarkably reduced the cross-sectional area of each muscle. The cross-sectional area of each muscle was quantified by ImageJ. The relative RNA and protein levels were normalized to those of the control β-actin. Error bars represent the mean ± SD of three biological replicates. *p < 0.05, **p < 0.01.