Journal: Frontiers in Veterinary Science
Article Title: miR-379-5p retards the proliferation and differentiation of goat skeletal muscle satellite cells by targeting LIN28B
doi: 10.3389/fvets.2025.1694160
Figure Lengend Snippet: miR-379-5p inhibits differentiation and promotes mitochondrial function in differentiating goat MuSCs. (A) Relative expression level of miR-379-5p in goat MuSCs transfected with NC or miR-379-5p mimics during the differentiation phase. (B) Relative mRNA levels of differentiation marker genes ( MyoD , MyoG , MyHC , Myf5 , and MEF2C ) in MuSCs transfected with NC or miR-379-5p mimics. (C) Western blot analysis of MyoD protein levels in differentiating MuSCs transfected with NC or miR-379-5p mimics. β-Tubulin was used as a loading control. (D) Representative immunofluorescence images of MyHC and DAPI in differentiating goat MuSCs after miR-379-5p overexpression. The right panel shows quantification of the fusion index. Scale bar = 50 μm. (E) Relative mRNA levels of mitochondrial marker genes ( CYCS , ATP5a1 , TFAM , and COX1 ) in differentiating MuSCs transfected with NC or miR-379-5p mimics. (F) Representative fluorescence images and quantification of mitochondrial membrane potential and ROS levels in differentiating MuSCs. Left: JC-1 staining reflects mitochondrial membrane potential. ROS levels were indicated by green fluorescence. Middle: Ratio of fluorescence intensity. Right: Quantification of ROS fluorescence intensity. Scale bar = 500 μm. Each experiment contained three biological replicates. * p < 0.05, ** p < 0.01.
Article Snippet: Primary antibodies included Pax7 (1:200, Santa Cruz Biotechnology, China), PCNA (1:200, Santa Cruz Biotechnology, China), β -Tubulin mouse mAb (1:5000, ZenBioScience, China), and MyoD polyclonal antibody (1:1000, Proteintech, United States).
Techniques: Expressing, Transfection, Marker, Western Blot, Control, Immunofluorescence, Over Expression, Fluorescence, Membrane, Staining