Figure 3.

Identification of key molecules associated with milk EVs‐mediated wound healing. A) Representative western blot of three (n = 3) independent experiments showing the expression levels of proteins in the Smad2/3 signaling pathway (Smad2/3, p‐Smad2, Smad7, and GAPDH). NIH‐3T3 cells were treated with PBS, EVs (100 µg mL⁻¹), or TGF‐β (10 ng mL⁻¹) and incubated for 24 h. B) Western blot quantification of Smad protein levels. C) Quantitative PCR analysis of the relative expression levels of miRNA‐21 in each EV group. D) Wound scratch migration assay in NIH‐3T3 cells (100 µg mL⁻¹: 5×10⁷ particles mL⁻¹) treated with anti‐miRNA‐21. All data are presented as mean ± SD of three independent experiments (*p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001 vs saline control, Serum EVs group; Dunnett's multiple comparisons test).