Af7584

For immunohistochemical analysis, mice were first anesthetized with ketamine (16 mg/kg body weight) and perfused transcardially with phosphate-buffered saline (PBS), followed by 4% paraformaldehyde (PFA) in PBS. The right hind limbs of the mice were removed from the skin and gastrocnemius muscle and immersed in 4% PFA overnight. The tibia and excess muscles were then removed, and the TA muscles were fixed for 3 h in 4% PFA, then dehydrated with 30% sucrose for 24 h. The TA muscles were then embedded in optimal cutting temperature solution (OCT), frozen, and sectioned at a thickness of 10 µm. The sections were blocked and permeabilized with 10% normal bovine serum and 0.2% 30%Triton X-100 in PBS for 2 h, then incubated overnight at 4 °C with primary antibodies. Subsequently, the sections were washed with PBS and stained with the Alexa-Fluor-488 or Alexa-Fluor-594-labeled goat anti-mouse or rabbit Ig secondary antibodies (diluted 1:500). For section immunofluorescence, the following antibodies were used: TMEM30A (rabbit polyclonal, ab217330, Abcam, USA), Pax7 (rabbit polyclonal, AF7584, Affinity, USA), Laminin (rabbit polyclonal, L9393, Sigma, USA), MYH3 (rabbit polyclonal, 22287-1-AP, Proteintech, USA), MYOD (rabbit polyclonal, 18943-1-AP, Proteintech, USA), and MYOG (rabbit polyclonal, bs-3550R, Bioss, USA).