Research Paper Volume 12, Issue 18 pp 18545—18560

MiR-520d-5p modulates chondrogenesis and chondrocyte metabolism through targeting HDAC1

Figure 2. miR-520d-5p affects the expression of HDAC1 in hMSCs during chondrogenesis. In these experiments, hMSCs were transfected with miR-520d-5p inhibitor (miR-520d-5p-IN) or miR-520d-5p mimics (miR-520d-5p-MI) at 0 day and then were induced to differentiate into chondrocytes for 21 days. All experiments were conducted at 21 days after chondrogenic induction. (A, B) Expressions of miR-520d-5p in hMSCs. (C) mRNA expressions of HDAC1 in hMSCs. (D) mRNA expressions of the chondrogenic markers AGGRECAN, COMP, COL2A1, and SOX9 and the hypertrophic markers COL10A1 and RUNX2 in hMSCs transfected with miR-520d-5p-IN at 0 days of chondrogenesis. (E) Protein expressions of HDAC1, SOX9, and COL2A1 in hMSCs transfected with miR-520d-5p-IN at 0 days of chondrogenesis. (F) mRNA expressions of the chondrogenic markers AGGRECAN, COMP, COL2A1, and SOX9 and the hypertrophic markers COL10A1 and RUNX2 in hMSCs transfected with miR-520d-5p-MI at 0 days of chondrogenesis. (G) Protein expressions of HDAC1, SOX9, and COL2A1 in hMSCs transfected with miR-520d-5p-MI at 0 days of chondrogenesis. (H) Immunohistochemistry assay for collagen type II in hMSCs transfected with miR-520d-5p-IN or miR-520d-5p-MI at 0 days of chondrogenesis (16× magnification). Scale bar = 100 μm. For each experiment, at least three replicates were available for the analysis. Data were expressed as mean ± standard deviation (SD). *P < 0.05; ** P < 0.01; *** P < 0.001.