Research Paper Volume 16, Issue 5 pp 4609—4630

Figure 2. Production and identification engineered exosomes containing miR-140-5p mimics or inhibitors. Preparation and identification of MPC-EVs. (A) illustrates the formation of myotubes post differentiation of C2C12 cells, shown via IF with red representing the heavy chain myosin (MYH) and blue for DAPI-stained nuclei (Scale bar in 200 μm). (B) depicts the preparation process of MPC-EVs. (C) presents the TEM results, showing the characteristic cup-shaped or semispherical morphology of the myogenic extracellular vesicles, in agreement with the expected bilayer membrane ultrastructure of extracellular vesicles. (D) shows WB results indicating positive reactions for the marker proteins TSG101, CD9, and CD63 in all types of MPC-EVs. (E) illustrates RT-PCR results (fold change), showing the miR-140-5p levels in MPC-Exo¹⁴⁰⁺ and MPC-Exo¹⁴⁰⁻, which are 1800 times and 1/3 of normal extracellular vesicles, respectively. MPC-Exo¹⁴⁰⁺-NC and MPC-Exo¹⁴⁰⁻-NC showed no difference from normal extracellular vesicles. Different letters between bars mean P ≤ 0.05 analyses followed by non-paired Student’s t-test. ^nsp > 0.05, ^*p < 0.05, ^**p < 0.01 and ^***p < 0.001 vs. MPC-Exo.