Research Paper Volume 9, Issue 3 pp 1012—1029

MicroRNA-434-3p regulates age-related apoptosis through eIF5A1 in the skeletal muscle

Figure 4. miR-434-3p protects apoptosis through eIF5A1. Myotubes were transfected with NS-mimetic, miR-434-3p mimetic with or without miR-434-3p antagomir or no transfection for 24 h followed by eight hours TPEN (100 μM) or Stps (1 μM) incubation. (A and B) Apoptosis (%) was measured by MTT assay in TPEN treated and non-treated myotubes (A) and the expression of eIF5A1 protein levels were determined by western blot (B). (C and D) Apoptosis (%) was measured by MTT assay in Stps treated and non-treated myotubes (C) and the expression of eIF5A1 protein levels were determined by western blot (D). (E) Myotubes were transfected with non-specific siRNA (NS-siRNA) or eIF5A1 siRNA for 24 h followed by eight hours TPEN (100 μM) or Stps (1 μM) incubation. Apoptosis (%) was measured by MTT assay. (F) miR-434-3p and eIF5A1 levels in young and aging GA muscles were determined by solution hybridization and western blot methods, respectively. U6 and GAPDH were served as loading controls for miR-434-3p and eIF5A1, respectively. Gel pictures are representative of three independent experiments. Each bar indicates mean ± SEM (n = 3). *, p < 0.05 vs. control; ϕ, p < 0.05 vs. NS-miR or NS-siRNA; δ, p < 0.05 vs. mimic.