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. 2017 Jan 27;25(3):715–727. doi: 10.1016/j.ymthe.2016.12.020

Figure 5.

Figure 5

miR-483-5p Directly Targets Matn3 in Chondrocytes during OA Development

(A) miR-483-5p aligned with the 3′ UTR of Matn3 mRNA. (B) ATDC5 cells were co-transfected with a reporter carrying a mutant or WT Matn3 3′ UTR along with miR-483-5p mimics or its control were analyzed by luciferase assay. (C) Western blot and real-time PCR analysis of Matn3 expression in ATDC5 cells transfected with miR-483-5p mimics or inhibitor for 48 hr. (D) Real-time PCR analysis of Col2a1, Ihh, and Runx2 mRNA levels in primary chondrocytes transfected with NC, miR-483-5p mimics, or miR-483-5p mimics plus Matn3 for 48 hr. (E) Immunohistochemical analysis of Matn3-positive cells in the tibial plateau of TG483 (n = 10) and control mice (n = 8) treated with 2 mg/mL Dox for 12 months. Scale bar, 20 μm. (F) H&E, safranin O, and fast green staining and immunostaining analysis of Matn3-, Ihh-, and Runx2-positive cells (brown) in the tibial plateau of DMM TG483 mice with or without intra-articular injection of Matn3-expressing lentivirus (n = 10). Red arrows indicate positively stained cells. Error bars represent the mean ± SEM. *p < 0.05; **p < 0.01; ns, non-significant (independent-sample t test for two groups).