Research Paper Volume 12, Issue 18 pp 18545—18560

Figure 4. miR-520d-5p regulates the chondrocyte metabolism through targeting HDAC1. (A, B) Expressions of miR-520d-5p and HDAC1 in PHCs treated with miR-520d-5p negative control (miR-520d-5p-NC) or miR-520d-5p mimics at 24 hours post-treatment, respectively. (C) mRNA expressions of the chondrogenic markers AGGRECAN, COMP, COL2A1, and SOX9 and the hypertrophic markers COL10A1 and RUNX2 in PHCs treated with miR-520d-5p negative control (miR-520d-5p-NC) or miR-520d-5p mimics at 24 hours post-treatment. (D, E) Expressions of miR-520d-5p and HDAC1 in PHCs treated with miR-520d-5p negative control (miR-520d-5p-NC) or miR-520d-5p inhibitors at 24 hours post-treatment, respectively. (F) mRNA expressions of the chondrogenic markers AGGRECAN, COMP, COL2A1, and SOX9 and the hypertrophic markers COL10A1 and RUNX2 in PHCs treated with miR-520d-5p negative control (miR-520d-5p-NC) or miR-520d-5p inhibitors at 24 hours post-treatment. (G–H) Expressions of miR-520d-5p and HDAC1 in OA chondrocytes treated with miR-520d-5p negative control (miR-520d-5p-NC) or miR-520d-5p mimics at 24 hours post-treatment, respectively. (I) mRNA expressions of the chondrogenic markers AGGRECAN, COMP, COL2A1, MMP-13, and SOX9 in OA chondrocytes treated with miR-520d-5p negative control (miR-520d-5p-NC) or miR-520d-5p mimics at 24 hours post-treatment. (J, K) Expressions of miR-520d-5p and HDAC1 in OA chondrocytes treated with miR-520d-5p negative control (miR-520d-5p-NC) or miR-520d-5p inhibitors at 24 hours post-treatment, respectively. (L) mRNA expressions of the chondrogenic markers AGGRECAN, COMP, COL2A1, MMP-13, and SOX9 in OA chondrocytes treated with miR-520d-5p negative control (miR-520d-5p-NC) or miR-520d-5p inhibitors at 24 hours post-treatment. (M) Acetylation of histone H3 in PHCs treated with miR-520d-5p negative control (miR-520d-5p-NC), miR-520d-5p inhibitors, or miR-520d-5p mimics at 48 hours post-treatment, respectively. (N) The putative or mutated miR-520d-5p binding site at 3'UTR of HDAC1. (O) Luciferase reporter assay in PHCs with wild-type or mutant 3’UTR in the presence of miR-520d-5p negative control (miR-520d-5p-NC), miR-520d-5p inhibitors, or miR-520d-5p mimics. 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.