Journal: Frontiers in Physiology

Article Title: Age-Related Exosomal and Endogenous Expression Patterns of miR-1, miR-133a, miR-133b, and miR-206 in Skeletal Muscles

doi: 10.3389/fphys.2021.708278

Figure Lengend Snippet: SRF and Cx43 production is inversely associated to the myomiR expression. The function of the four myomiRs in the five skeletal muscles was further explored by evaluating the production levels of two of their established targets, SRF and Cx43. (A–E) Fold change (FC) levels obtained from western blot analysis of SRF and Cx43 across different ages and compared the 2-week levels are shown. SRF was determined to be inversely correlated to the endogenous expression of miR-1, miR-133a, and miR-133b for all five skeletal muscles. Cx43 was found to be inversely correlated to the endogenous expression profile of miR-206. The glycolytic muscles (A) EDL, (C) TA, (D) gastrocnemius, and (E) quadriceps were shown to express increasing levels of Cx43 as the mice age, while (B) the oxidative soleus muscle was identified to express decreasing Cx43 levels with muscle development. Representative western blot images are shown for each skeletal muscle. GAPDH was served as normalization control in three western blot analyses ( n = 3). Data on graphs represent the mean values ± SEM.

Article Snippet: The next day, membranes were washed thrice and incubated with anti-mouse IgG or anti-rabbit IgG secondary antibodies conjugated to HRP (1:5,000 dilution; Santa Cruz Biotechnology) for 2 h. Proteins were visualized using UVP BioSpectrum ® 810 Imaging System or Vilber Fusion Solo S. To determine changes in protein abundances in skeletal muscle tissues across time, SRF and Cx43 concentration levels were normalized over the internal GAPDH control.

Techniques: Expressing, Western Blot