Fig. 5

Fam3a is secreted by myogenic cells. a, b Immunofluorescence analysis of C2C12 myogenic cells transfected with a construct to overexpress Fam3a-Myc-Flag. GM-130 is a cis-Golgi marker. KDEL is an endoplasmic reticulum marker. Purple or yellow colors mean colocalization. When indicated, cells were treated with Monensin (200 μM) for 4 h. Scale bars 20 μm. c, d Western blot analysis of non-transfected C2C12 myoblasts and C2C12 cells transfected with constructs to overexpress green fluorescent protein (GFP) or Fam3a-Myc-Flag. When indicated, cells were treated with Monensin (200 μM) for 4 h (n = 3 independent experiments). e Western blot analysis of the media from C2C12 cells transfected with a construct to overexpress Fam3a-Myc-Flag. f Fam3a mRNA abundance per cell in freshly isolated muscle stem cells (MuSCs), macrophages (MPs), fibroadipogenic progenitors (FAPs), and myofibers from C57BL6/J P15 pups (n = 3 biologically independent samples; 3 pups were pulled per sample) and 3-month-old adult male mice (n = 3 animals). g Fam3a mRNA abundance per population in freshly isolated MuSCs, MPs, and FAPs from uninjured, 3 dpi, and 7 dpi 3-month-old male mice (n = 3 animals). Data are represented as mean ± SEM (Student’s t test; *p < 0.05, **p < 0.01, ****p < 0.0001)