Fig. 2

miR-101-3p mimic promoted HAVICs calcification and inhibition of miR-101-3p protected HAVICs from calcification in CM. Alizarin red staining (A–F) of HAVICs cultured in osteogenic-conditioned media (10 mM βGP, 5 mM CaCl₂) for 14 days after transfected with miR-101-3p mimic, inhibitor and negative control. G Quantification of alizarin red staining, absorbance normalized by protein concentration. *P < 0.05, ***P < 0.001 and ****P < 0.0001 vs Control; ⁺⁺⁺⁺P < 0.0001 vs Mimic; ^P < 0.05, ^^^^P < 0.0001 vs Control, CM; ^####P < 0.0001 vs Inhibitor. H Western blot analysis of Fetuin A protein expression in HAVICs cultured in CM for 48 h after miR-101 mimic/inhibitor transfection. β-actin serves as internal loading control. I, J Quantification of Fetuin A protein expression normalized by β-actin. Data = means ± SEM. ****P < 0.0001 vs Control; ^P < 0.05 and ^^^^P < 0.0001 vs Mimic or Inhibitor groups; ⁺⁺⁺⁺P < 0.0001 vs Control, CM. n = 3 repeats